CN107207521A

CN107207521A - It is used as the substituted bridging urea analog of Sirtuin conditioning agent

Info

Publication number: CN107207521A
Application number: CN201580073782.4A
Authority: CN
Inventors: J.L.埃利斯; K.A.埃文斯; R.M.福克斯; W.H.米勒; M.A.塞费尔德
Original assignee: GlaxoSmithKline Intellectual Property No 2 Ltd
Current assignee: GlaxoSmithKline Intellectual Property No 2 Ltd
Priority date: 2014-11-19
Filing date: 2015-11-19
Publication date: 2017-09-26
Also published as: WO2016079712A1; CA2968029A1; KR20170087907A; WO2016079710A1; WO2016079711A1; KR20170083133A; US10072011B2; ES2823748T3; AU2015348943A1; CR20170209A; BR112017010599A2; EP3220918A1; EP3221316A1; CN107108627A; SG11201703824QA; IL252252A0; EP3221322A1; CN107207520A; CO2017004994A2; BR112017010595A2

Abstract

The present invention relates to the new substituted bridging urea analog compounds of formula (I) or its pharmaceutically acceptable salt, corresponding pharmaceutical composition, prepare and using the method for the compound, the compound individually or with combination with other therapeutic agents is used to improve cell survival as Sirtuin conditioning agent, and a variety of diseases and obstacle for the treatment of and/or prevention wide scope, the disease and obstacle include but is not limited to, for example, the disease relevant with aging or stress reaction or obstacle, diabetes, obesity, neurodegenerative disease, angiocardiopathy, blood coagulation disorder, inflammation, cancer and/or flush, and benefit from mitochondrial active increased disease or obstacle.

Description

It is used as the substituted bridging urea analog of Sirtuin conditioning agent

Technical field

Generally, the present invention relates to the substituted bridging urea analog compounds of formula (I) to (V), its corresponding analog Or derivative, or its pharmaceutically acceptable salt, corresponding pharmaceutical composition, the method for preparing the compound, the chemical combination Thing individually or with combination with other therapeutic agents as Sirtuin (Sirtuin) conditioning agent is used for following method and purposes： Cell survival, and treatment and/or a variety of diseases and obstacle of preventing wide scope are improved, the disease and obstacle include but do not limited In for example, the disease relevant with aging or stress reaction or obstacle, diabetes, obesity, neurodegenerative disease, cardiovascular disease Disease, blood coagulation disorder, inflammation, cancer and/or flush, and benefit from mitochondrial active increased disease or obstacle.

Background technology

Silent message conditioning agent (Silent Information Regulator, SIR) gene family, which is represented, is present in model Enclose one group of highly conserved gene into the genome of Eukaryotic organism from archeobacteria (archaebacteria).Compile The SIR albumen of code is related to the various various process repaired from the regulation of gene silencing to DNA.Well-characterized gene in this family For Saccharomyces Cerevisiae in S IR2 (S.cerevisiae SIR2), it, which involves, makes containing regulation yeast mating type, telomere position effect and thin The HM locus silences of the information of born of the same parents' aging.Yeast Sir2 albumen belongs to the family of histone deacetylase.By SIR genes man The albumen of race member coding is shown：There is high degree of sequence to guard in the Core domain of 250 amino acid.In Salmonella typhimurium Sir2 homologues CobB in bacterium (Salmonella typhimurium) play NAD (NADH)-according to Rely the effect of property ADP- ribosyltransferases.

Sir2 albumen is to use NAD as the group iii deacetylase of cosubstrate.Different from other deacetylations Enzyme, many in these is related to gene silencing, and Sir2 is (such as bent to I classes and class ii histone deacetylase inhibitors Ancient ablastins A (trichostatin A, TSA)) do not have sensitiveness.

Combined closely by the deacetylated and NAD hydrolysises of the Sir2 acetyl-lysines carried out, produce nicotinoyl Amine and new acetyl group-ADP ribose compounds.Sir2 NAD- dependences deacetylase activity is for can be by its biology It is required to act on for acting on this function of combining with the cell metabolism in yeast.Mammal Sir2 homologues have NAD- dependence histone deacetylase activities.

Biochemical research is it has been shown that Sir2 can be easily by histone H 3 and H4 amino terminal tails deacetylation Change, result in 2 '/3 '-O- acetyl group-ADP- ribose (OAADPR) and niacinamide.The bacterial strain of additional copy with SIR2 shows Show the increase of rDNA silences and life 30%.Have also shown that, Caenorhabditis elegans (C.elegans) SIR2 homologues And the additional copy of drosophila (D.melanogaster) dSir2 genes can extend the life-span of those organisms (sir-2.1).This is dark The SIR2- dependences regulation approach shown for aging (aging) occurs in early stage of evolving and has been quite conservative.It is existing It is modern, it is believed that Sir2 genes have entered to turn to the health of enhancing organism with that stress resist, to increase the chance of its adverse circumstance survival.

There are 7 kinds of Sir2 samples genes (SIRT1-SIRT7) in people, their shared Sir2 conserved catalytic domain.SIRT1 For with the nucleoprotein with Sir2 top sequence similarities.SIRT1 adjusts various kinds of cell target spot by deacetylated effect, Including caused by tumor suppressor p 53, cellular signal transduction factor NF-kB and FOXO transcription factor.

SIRT3 is SIRT1 homologue, and it has conservative in prokaryotes and eucaryote.SIRT3 albumen passes through position In N- ends unique domain and targeted to mitochondrial cristae (cristae).SIRT3 has NAD⁺- dependence protein is deacetylated Base enzymatic activity, and generally express, particularly in the tissue of metabolic activity.After mitochondria is transferred to, it is believed that SIRT3 is by line grain Body matrix processing peptidase (MPP) fragments into smaller activity form.

Heat limitation (caloric restriction) is promoted the health of mammal and the fact that extend the life-span behaved Known more than 70 years.The yeast life-span (life-span of such as metazoa) limits the intervention of (such as low glucose) also by similar heat To increase.It was found that the yeast for lacking SIR2 genes is not lived longer with flies when heat is limited, this is SIR2 gene mediateds The effect good for health of restricted calorie diet provides evidence.Moreover, reduction yeast glucose-response cAMP (adenosine 3', 5'- monophosphates) mutation of-dependence (PKA) pathway activities extends the life-span of wild-type cell, but in saltant type sir2 bacterial strains In will not then extend, this prove SIR2 be probably heat limit approach crucial downstream component.

Except treatment potentiality, the structure of SIRT1 activity and biophysics research and through small molecule Sirtuin (sirtuin) activation of conditioning agent by for promote the understanding to the biological function of Sirtuin, further appreciate that it is heavy The experiment for identifying new Sirtuin conditioning agent is researched and developed in the mechanism of action of silent regulatory protein activation and help.

Present invention seek to address that these and other problems run into this area.

The content of the invention

Generally, the present invention relates to the substituted bridging urea analog compounds of formula (I) to (V), its corresponding analog Or derivative, or its pharmaceutically acceptable salt, corresponding pharmaceutical composition, the method for preparing the compound, and it is described Compound individually or with combination with other therapeutic agents as Sirtuin conditioning agent is used for following purposes：Improve the cell longevity Life, and treatment and/or a variety of diseases and obstacle of preventing wide scope, the disease and obstacle include but is not limited to, for example, with Aging or the relevant disease of stress reaction or obstacle, diabetes, obesity, neurodegenerative disease, angiocardiopathy, blood clotting Obstacle, inflammation, cancer and/or flush, and benefit from mitochondrial active increased disease or obstacle.

In particular it relates to which the noval chemical compound of formula (I) to (V), corresponding analog are (that is, in R²Position has hydrogen Substituted analog) and respectively comprising formula (I) to (V) compound corresponding pharmaceutical composition.

The invention further relates to prepare formula (I) respectively to the compound and corresponding analog of (V) (that is, in R²Position has Hydrogen substitution analog) method.

The invention further relates to treated using application-defined Sirtuin regulation immunomodulator compounds, prevented wide scope A variety of diseases and obstacle or a variety of diseases and the therapy of obstacle for wide scope in method and purposes, the disease and Obstacle includes but is not limited to, for example, the disease relevant with aging or stress reaction or obstacle, diabetes, obesity, neurodegeneration Disease, angiocardiopathy, blood coagulation disorder, inflammation, cancer and/or flush, and to benefit from mitochondrial activity increased Disease or obstacle, it is further selected from or included but is not limited to, psoriasis, atopic dermatitis, acne, brandy nose, inflammatory bowel disease, bone Matter osteoporosis, septicemia, arthritis, COPD, systemic loupus erythematosus and ophthalmic inflammation.

Detailed description of the invention

Compound

International patent application WO09/061879 discloses the regulation silence of formula (I) in (international filing date on May 13rd, 2014) The substituted bridging urea and related analogs compound of regulatory protein：

Or

Its pharmaceutically acceptable salt, corresponding pharmaceutical composition, the combination with other therapeutic agents, its preparation method and For following method and purposes：Improve cell survival, and treatment and/or a variety of diseases and obstacle of preventing wide scope, institute Stating disease and obstacle includes, for example, the disease relevant with aging or stress reaction or obstacle, diabetes, obesity, neurodegeneration Disease, angiocardiopathy, blood coagulation disorder, inflammation, cancer and/or flush, and to benefit from mitochondrial activity increased Disease or obstacle.

In one aspect, structure formula (I) of the present invention to the new regulation Sirtuin of (V) compound, respective Corresponding analog is (that is, in R²Position has the analog that hydrogen replaces), as described in detail.

In one aspect, the present invention relates to formula (I) compound：

Wherein：

X₁Or X₂It is independently selected from-N or-C；

R¹Heterocyclic radical, aryl, heteroaryl ,-C (O) R replaced for hydrogen, halogen ,-CN, carbocylic radical, heterocyclic radical ,-N-_aOr-C (O)-NR_bR_c；

R²For halogen, straight or branched C₁-C₆Alkyl, straight or branched-C₁-C₆Haloalkyl, or-C (O)-NR_bR_c；

R³For hydrogen, halogen ,-hydroxyl, straight or branched C₁-C₆Alkyl, or straight or branched-C₁-C₆Haloalkyl；

R⁴For hydrogen or-C (O) NR_bR_c；

Wherein：

Work as X₂When being-N, R₂It is not present；Or

Work as X₂When being-C, R₂As defined above；

Each R as defined above¹、R²、R³Or R⁴Optionally further it is selected from following one or more substituents substitution：Hydrogen, Halogen ,-OH ,-(CH₂)_xOH、-C≡N、-NR_dR_e, straight or branched C₁-C₆Alkyl, straight or branched-C₁-C₆It is haloalkyl, straight Chain or side chain C₁-C₆Alkoxy, straight or branched C₁-C₆Halogenated alkoxy ,-O- straight or brancheds-C₁-C₆Haloalkyl ,-C₁-C₆ Cycloalkyl ,-(CH₂)_x- cycloalkyl, heterocyclic radical, aryl ,-heteroaryl ,-(CH₂)_x- heteroaryl ,-O- (CH₂)_xCH(OH)CH₂ , or-C (O) OR (OH)_f；

Each R as defined above_a、R_b、R_c、R_d、R_eOr R_fIt is independently selected from hydrogen, straight or branched C₁-C₆Alkyl, straight or branched- C₁-C₆Haloalkyl ,-C₁-C₆- cycloalkyl ,-(CH₂)_xC₁-C₆- cycloalkyl, heterocyclic radical ,-N- heterocyclic radicals, aryl, heteroaryl, or- (CH₂)_xHeteroaryl ,-(CHR_g)_xHeteroaryl；

Wherein：

R_gFor straight or branched C₁-C₆Alkyl, straight or branched-C₁-C₆Haloalkyl；

Each R as defined above_a、R_b、R_c、R_d、R_eOr R_fOptionally following one or more substituents are further selected to take Generation：Hydrogen, halogen ,-OH ,-C ≡ N, straight or branched C₁-C₆Alkyl, straight or branched-C₁-C₆Haloalkyl, straight or branched C₁- C₆Alkoxy ,-O- straight or brancheds-C₁-C₆Haloalkyl ,-C₁-C₆Cycloalkyl, carbocylic radical ,-(CH₂)_x- carbocylic radical ,-heterocycle Base ,-O- heterocyclic radicals, aryl ,-heteroaryl ,-(CH₂)_x- heteroaryl ,-O- (CH₂)_xCH(OH)CH₂(OH)、-(CH₂)_x- OH, or-C (O)-OH；

M is integer 1 to 3；

N is selected from 1 to 3 integer；

X is 0 or integer 1 to 6；Or

Its pharmaceutically acceptable salt.

On the other hand, the present invention relates to the compounds of this invention such as defined in the whole text above and such as the application, i.e. structural formula The compound of (I) to (V), respectively analog are (that is, in R²Position has the analog that hydrogen replaces), its conditional It is：

As n=1,And

As n=3,

On the other hand, the present invention relates to the compounds of this invention, wherein R²It is C (O)-NR_bR_c；Wherein R_bAnd R_cAs above and As the application is defined in the whole text.

On the other hand, the present invention relates to formula (I) compound, wherein：

M is 1；

N is 2 or 3；And

R⁴It is hydrogen.

M is 1；

N is 2 or 3；And

R⁴It is-C (O) NR_bR_c, wherein each R_bAnd R_cAs defined above.

M is 1；

N is 2 or 3；

R¹Heterocyclic radical, aryl or the heteroaryl replaced for hydrogen, halogen ,-CN, carbocylic radical, heterocyclic radical ,-N-；And

R⁴It is-C (O) NR_bR_c, wherein R_bAnd R_cAs defined in following claims 1.

In one aspect, the present invention relates to formula (II) compound：

Wherein：

X₁Or X₂It is independently selected from-N or-C；

R¹Heterocyclic radical, aryl or the heteroaryl replaced for hydrogen, halogen ,-CN, carbocylic radical, heterocyclic radical ,-N-；

R⁴For hydrogen or-C (O) NR_bR_c；

Wherein：

Work as X₂When being-N, R₂It is not present；Or

Work as X₂When being-C, R₂As defined above；

Wherein：

Each R as defined above_a、R_b、R_c、R_d、R_eOr R_fOptionally following one or more substituents are further selected to take Generation：Hydrogen, halogen ,-OH ,-C ≡ N, straight or branched C₁-C₆Alkyl, straight or branched-C₁-C₆Haloalkyl, straight or branched C₁- C₆Alkoxy ,-O- straight or brancheds-C₁-C₆Haloalkyl ,-C₁-C₆Cycloalkyl, carbocylic radical ,-(CH₂)_x- carbocylic radical ,-heterocycle Base ,-O- heterocyclylalkyls ,-heteroaryl ,-(CH₂)_x- heteroaryl ,-O- (CH₂)_xCH(OH)CH₂(OH)、-(CH₂)_x- OH, or-C (O)-OH；

M is integer 1 to 3；

N is selected from 1 to 3 integer；

X is 0 or integer 1 to 6；Or

Its is pharmaceutically acceptable

On the other hand, the present invention relates to the compounds of this invention such as defined in the whole text with the application above (that is, structural formula The compound of (I) to (V), respectively analog is (that is, in R²Position has the analog that hydrogen replaces), its conditional It is：

As n=1,And

As n=3,

On the other hand, the present invention relates to the compounds of this invention, wherein R²It is C (O)-NR_bR_c；Wherein R_bAnd R_cAs above and As the application defines in the whole text

M is 1；

N is 2 or 3；And

R⁴It is hydrogen.

M is 1；

N is 2 or 3；And

R⁴It is-C (O) NR_bR_c, wherein each R_bAnd R_cAs defined above.

On the other hand, the present invention relates to formula (III) compound：

Wherein：

X₁Or X₂It is independently selected from-N or-C；

Wherein：

Work as X₂When being-N, R₂It is not present；Or

Work as X₂When being-C, R₂As defined above；

Each R⁵And R⁶It is independently selected from hydrogen, straight or branched C₁-C₆Alkyl, straight or branched-C₁-C₆Haloalkyl ,-C₁-C₆Ring Alkyl ,-(CH₂)_xC₁-C₆Cycloalkyl, heterocyclic radical ,-N- heterocyclic radicals, aryl, heteroaryl, or-(CH₂)_xHeteroaryl ,-(CHR_g)_xIt is miscellaneous Aryl；

Wherein：

Each R as defined above¹、R²、R³、R⁵And R⁶Optionally further it is selected from following one or more substituents substitution： Hydrogen, halogen ,-OH ,-(CH₂)_xOH、-C≡N、-NR_dR_e, straight or branched C₁-C₆Alkyl, straight or branched-C₁-C₆Haloalkyl, Straight or branched C₁-C₆Alkoxy, straight or branched C₁-C₆Halogenated alkoxy ,-O- straight or brancheds-C₁-C₆Haloalkyl ,- C₁-C₆Cycloalkyl ,-(CH₂)_x- cycloalkyl, heterocyclic radical, aryl ,-heteroaryl ,-(CH₂)_x- heteroaryl ,-O- (CH₂)_xCH(OH)CH₂ , or-C (O) OR (OH)_f；

Each R as defined above_a、R_b、R_c、R_d、R_e、R_fOr R_gIt is independently selected from hydrogen, straight or branched C₁-C₆Alkyl, straight chain or branch Chain-C₁-C₆Haloalkyl ,-C₁-C₆- cycloalkyl ,-(CH₂)_xC₁-C₆- cycloalkyl, heterocyclic radical ,-N- heterocyclic radicals, aryl, heteroaryl, Or-(CH₂)_xHeteroaryl；

Wherein：

Each R as defined above_a、R_b、R_c、R_d、R_e、R_fOr R_gOptionally further it is selected from following one or more substituents Substitution：Hydrogen, halogen ,-OH ,-(CH₂)_xOH ,-C ≡ N, NR_hR_i, straight or branched C₁-C₆Alkyl, straight or branched-C₁-C₆Halo Alkyl, straight or branched C₁-C₆Alkoxy, straight or branched-C₁-C₆Halogenated alkoxy ,-C₁-C₆Cycloalkyl ,-(CH₂)_x- cycloalkanes Base, heterocyclic radical ,-heterocyclic radical ,-O- heterocyclic radicals, aryl ,-heteroaryl ,-(CH₂)_x- heteroaryl ,-O- (CH₂)_xCH(OH)CH₂ (OH)、-(CH₂)_x- OH, or-C (O) OR_j；

Wherein：

Each R_h、R_iAnd R_jIt is independently selected from hydrogen, straight or branched C₁-C₆Alkyl or straight or branched-C₁-C₆Haloalkyl；

M is integer 1 to 3；

N is selected from 2 to 3 integer；

X is 0 or integer 1 to 6；Or

Its pharmaceutically acceptable salt.

On the other hand, the present invention relates to the compounds of this invention, wherein n is 2 or 3 and m is 1.

On the other hand, the present invention relates to formula (IV) compound：

Wherein：

Each R as defined above_a、R_b、R_c、R_d、R_e、R_fOr R_gOptionally further it is selected from following one or more substituents Substitution：Hydrogen, halogen ,-OH ,-(CH₂)_xOH ,-C ≡ N ,-NR_hR_i, straight or branched C₁-C₆Alkyl, straight or branched-C₁-C₆Halo Alkyl, straight or branched C₁-C₆Alkoxy, straight or branched-C₁-C₆Halogenated alkoxy ,-C₁-C₆Cycloalkyl ,-(CH₂)_x- cycloalkanes Base, heterocyclic radical ,-heterocyclic radical ,-O- heterocyclic radicals, aryl ,-heteroaryl ,-(CH₂)_x- heteroaryl ,-O- (CH₂)_xCH(OH)CH₂ (OH)、-(CH₂)_x- OH, or-C (O) OR_j；

Wherein：

M is integer 1 to 3；

N is selected from 2 to 3 integer；

X is 0 or integer 1 to 6；Or

Its pharmaceutically acceptable salt.

On the other hand, the present invention relates to formula (V) compound：

Wherein：

M is integer 1 to 3；

N is selected from 2 to 3 integer；

X is 0 or integer 1 to 6；Or

Its pharmaceutically acceptable salt.

On the other hand, the present invention relates separately to formula (I) to (V) compound, wherein R¹It is selected from：

On the other hand, the present invention relates separately to formula (I) to (V) compound, wherein R⁴It is selected from：

On the other hand, the present invention relates to the compound defined in the application table 1, page 684 is originated in：

On the other hand, the present invention relates to compound, it includes, but are not limited to the chemical combination defined in the table that is listed herein below Thing：

Table 1

On the other hand, the present invention relates to compound, it is corresponding analog or derivative of the invention (that is, in R² Position has the analog that hydrogen replaces)：

The non-meta substituted pyridine compounds of table 2- (wherein, ISOMER=isomers)

Term and definition

Part 1

Some compounds of the present invention can exist with specific geometrical isomerism or stereoisomeric forms in any ratio.The present invention includes institute The such compound having, it is including cis-and trans-isomer, (R)-and (S)-enantiomter, diastereoisomer, (D)- Isomers, (L)-isomers, its racemic mixture, and their other mixtures, as fallen within the scope of the present invention. Other asymmetric carbon atoms may be present in substituent (such as alkyl).All such isomers and its mixture are included in In the present invention.

Compound specifically described herein and its salt can also accordingly hydrate (for example, semihydrate, monohydrate, two Hydrate, trihydrate, tetrahydrate) or solvate presence.Suitable solvent for preparing solvate and hydrate is led to It can often be selected by those skilled in the art.

The compound and its salt can exist in amorphous or crystallization (including eutectic and polymorphic) form.

The compound of the regulation Sirtuin of the present invention advantageously adjusts level and/or the work of Sirtuin Property, the especially deacetylase activity of Sirtuin.

Individually or in addition to above-mentioned property, the compound of some regulation Sirtuins of the invention is effectively being adjusted Do not have one during the concentration of the deacetylation activity of Sirtuin (for example, SIRT1 and/or SIRT3 albumen) substantially Or multiple following activity：Suppress PI3- kinases, suppress alditol reductase (aldoreductase), suppress EGFR-TK, turn a work Change EGFR EGFR-TKs, coronary artery expansion or Antispasmodic activity.

" alkyl " or " alkane " is fully saturated straight chain or side chain non-aromatic alkyl.Typically, straight chain or branch The alkyl of chain has 1 to about 20 carbon atom, preferably with 1 to about 10 carbon atom, unless otherwise indicated.The sum of straight chain The example of the alkyl of side chain includes methyl, ethyl, n-propyl, isopropyl, normal-butyl, sec-butyl, the tert-butyl group, amyl group, hexyl, heptan Base and octyl group.C₁-C₄Straight chain or side chain alkyl is also referred to as " low alkyl group ".

In any foregoing embodiments, C₁-C₄Alkoxy-substituted group may include one or more alkoxy substitutions Base, such as one, two or three methoxyl group, or methoxyl group and ethyoxyl, for example.Exemplary C₁-C₄Alkoxy substituent bag Include methoxyl group, ethyoxyl, isopropoxy and tert-butoxy.

In any foregoing embodiments, hydroxyl-substituted group may include one or more hydroxyl substituents, such as two Individual or three oh groups.

" halogen " refers to F, Cl, Br or I.

" halogen-substitution " or " halo " represents that one or more hydrogen are replaced by F, Cl, Br or I.

In one aspect, term haloalkyl is defined as one or moreHydrogenAtom is replaced by halogen atomAlkylIt is residual Base.In any foregoing embodiments, " halogen-substituted " group includes a halogenic substituent to up to perhalogeno and replaced Base.The C of exemplary halogen-substituted₁-C₄Alkyl includes CFH₂、CClH₂、CBrH₂、CF₂H、CCl₂H、CBr₂H、CF₃、CCl₃、 CBr₃、CH₂CH₂F、CH₂CH₂Cl、CH₂CH₂Br、CH₂CHF₂、CHFCH₃、CHClCH₃、CHBrCH₃、CF₂CHF₂、CF₂CHCl₂、 CF₂CHBr₂、CH(CF₃)₂With C (CF₃)₃.The C of perhalogeno₁-C₄Alkyl is for example including CF₃、CCl₃、CBr₃、CF₂CF₃、CCl₂CF₃With CBr₂CF₃。

Term " alkenyl " (" alkene ") and " alkynyl " (" alkynes ") are referred in length and may replaced and above-mentioned alkane Undersaturated aliphatic group as base class, but at least one each self-contained double bond or three key.

In any foregoing embodiments, " carbocyclic ring " can refer to monocycle carbocyclic ring embodiment and/or polycyclic carbocyclic ring embodiment party Case, such as fusion, bridging or bicyclic carbocyclic embodiment.It is real that " carbocyclic ring " group of the present invention can further refer to aromatic carbocyclic Scheme and/or non-aromatic carbocyclic ring embodiment are applied, or in the case of polycyclic embodiment, with one or more aromatic rings And/or the carbocyclic ring of both one or more non-aromatic rings.Polycyclic carbocyclic ring embodiment can be that bicyclic, fused rings or bridging are bicyclic. Nonrestrictive exemplary carbocyclic ring include phenyl, hexamethylene, pentamethylene or cyclohexene, amantadine, pentamethylene, hexamethylene, Bicyclic [2.2.1] heptane, 1,5- cyclo-octadiene, 1,2,3,4- naphthanes, bicyclic [4.2.0] octyl- 3- alkene, naphthalene, adamantane, decahydro Change naphthalene, naphthalene, 1,2,3,4- naphthanes, norcamphane, decahydronaphthalenes, spiropentane, Memantine hydrochloride (memantine), pyrrole Pai Lideng (biperiden), Rimantadine (rimantadine), camphor, cholesterine, 4- phenylcyclohexanols, bicyclic [4.2.0] octane, U.S. Amantadine and 4,5,6,7- tetrahydrochysene -1H- indenes and bicyclic [4.1.0] hept- 3- alkene.

In any foregoing embodiments, " heterocycle " group can refer to monocyclic heterocycles embodiment and/or polycyclic heterocycle is real Apply scheme, such as fusion, bridging or bicyclic heterocycle embodiment." heterocycle " group of the present invention can further refer to fragrance Heterocyclic ring embodiment and/or nonaromatic heterocycles embodiment, or in the case of polycyclic embodiment, with one or more virtues The heterocycle of both fragrant ring and/or one or more non-aromatic rings.Polycyclic heterocyclic ring embodiment can be bicyclic, fused rings or bridging Ring.Nonrestrictive Exemplary heterocyclic includes pyridine, pyrrolidines, piperidines, piperazine, pyrrolidines, morpholine, pyrimidine, benzofuran, Yin Diindyl, quinoline, lactone, lactams, benzodiazepineIndoles, quinoline, purine, adenine, guanine, 4,5,6,7- tetrahydrochysene benzene And [d] thiazole, urotropine (hexamine) and methenamine (methenamine).

" alkenyl " refers to the member carbon atoms with specified quantity and has one or more carbon-to-carbon double bonds in chain Aliphatic unsaturated hydrocarbon.For example, C2-C6 alkenyls refer to the alkenyl with 2 to 6 member carbon atoms.In certain embodiments, alkenyl There is a carbon-to-carbon double bond in chain.In other embodiments, alkenyl has more than one carbon-to-carbon double bond in chain.Alkenyl Optionally it can be replaced by one or more substituents as defined herein.Alkenyl can be straight or branched.Representational branch Alkenyl has one, two or three branches.Alkenyl includes vinyl, acrylic, cyclobutenyl, pentenyl and hexenyl.

" alkoxy " refers to the moieties (- O-C1-C6 that i.e. wherein C1-C6 is defined herein connected by oxygen bridge Alkyl).The example of these groups includes methoxyl group, ethyoxyl, propoxyl group, butoxy, amoxy and hexyloxy.

" alkynyl " refers to there is one or more keys of carbon-to-carbon three with the member carbon atoms specified number and in chain Aliphatic unsaturated hydrocarbon.For example, C2-C6 alkynyls refer to the alkynyl with 2 to 6 member atoms.In certain embodiments, alkynyl exists There is a key of carbon-to-carbon three in chain.In other embodiments, alkynyl has the more than one key of carbon-to-carbon three in chain.In order to clear Chu Qijian, has the aliphatic unsaturated hydrocarbon of one or more keys of carbon-to-carbon three in chain and has one or more carbon-to-carbons double in chain The aliphatic unsaturated hydrocarbon of key is referred to as alkynyl.Alkynyl optionally can be replaced by one or more substituents as defined herein.Represent Property branch alkynyl have one, two or three branches.Alkynyl includes acetenyl, propinyl, butynyl, pentynyl and hexin Base.

Term " aromatic carbocyclic " refers to including the armaticity hydrocarbon ring system of at least one aromatic rings.The ring can be fused to or It is connected on other aromatic carbocyclics or on non-aromatic carbocyclic ring.The example of aromatic carbon ring group includes carbocyclic aromatic radical such as benzene Base, naphthyl and anthryl.

" azabicyclo " refers to including the bicyclic molecule of nitrogen-atoms in ring skeleton.Two bicyclic rings can be at two Condensed at bonded atom mutually, such as indoles is condensed, such as azabicyclo [2.2.1] heptane across a series of atom, or Connected at single atom, for example, loop coil.

" bicyclic " or " bicyclic " refers to bicyclic ring system, wherein between the two rings share one, two or three or More atoms.It is bicyclic including condensed-bicyclic, the adjacent atom of two of which is each shared by the two rings, for example, decahydro Change naphthalene, indoles.Bicyclic also bicyclic including spiral shell, two of which ring shares an atom, for example, spiral shell [2.2] pentane, 1- oxa-s -6- Azaspiro [3.4] octane.Bicyclic also bicyclic including bridging, wherein at least three atom is shared by two rings, for example, drop camphane Alkane.

" bridging is bicyclic " compound is bicyclic system, and wherein at least three atom is shared by two rings of the system, i.e., They include at least one bridge in one or more atoms of two bridgehead atoms of connection.The azabicyclo of bridging is referred to The bicyclic molecule of bridging comprising nitrogen-atoms at least one ring.

Term " Boc " refers to t-butyloxycarbonyl (conventional amine protecting group).

Terms used herein " carbocyclic ring " and " carbocyclic ring " refer to saturated or unsaturated ring, the wherein ring each Atom is carbon.Term carbocyclic ring includes both aromatic carbocyclic and non-aromatic carbocyclic ring.Non-aromatic carbocyclic ring, which includes cycloalkanes hydrocarbon ring, (wherein to be owned Carbon atom be saturation) and both cyclenes hydrocarbon ring (its include at least one double bond)." carbocyclic ring " includes 5-7 unit monocycles and 8-12 Membered bicyclic.Each bicyclic carbocyclic ring may be selected from non-aromatic ring and aromatic rings.Carbocyclic ring include bicyclic molecule, one of them, two Or three or more atoms are shared by the two rings.Term " fused iso " refers to that wherein each ring is shared with another ring The bicyclic carbocyclic of two adjacent atoms.Each ring of fused iso may be selected from non-aromatic ring and aromatic rings.In exemplary reality Apply in scheme, aromatic rings (such as phenyl) can be fused to non-aromatic ring or aromatic rings, for example, hexamethylene, pentamethylene or hexamethylene Alkene.Non-aromatic and aromatic bicyclic any combination (as long as valence link permission) is included in the definition of carbocyclic ring.Exemplary " carbocyclic ring " Including pentamethylene, hexamethylene, bicyclic [2.2.1] heptane, 1,5- cyclo-octadiene, 1,2,3,4- naphthanes, bicyclic [4.2.0] octyl- 3- alkene, naphthalene and adamantane.Exemplary fused iso is pungent including decahydronaphthalenes, naphthalene, 1,2,3,4- naphthanes, bicyclic [4.2.0] Alkane, 4,5,6,7- tetrahydrochysene -1H- indenes and bicyclic [4.1.0] hept- 3- alkene." carbocyclic ring " can have hydrogen former at any one or more The position of son is substituted.

" cycloalkyl " is the ring-type hydrocarbon ring with the member carbon atoms specified number, and it is fully saturated (non-aromatic). Generally, group of naphthene base has 3 to about 10 carbon atoms, more typically 3 to 8 carbon atoms, unless otherwise defined.Cycloalkyl is Single ring systems.For example, C3-C6 cycloalkyl refers to the cycloalkyl with 3 to 6 member atoms.Cycloalkyl can be optionally by one Individual or multiple substituent substitutions as defined herein.Cycloalkyl includes cyclopropyl, cyclobutyl, cyclopenta and cyclohexyl.

" cycloalkenyl group " is the ring-type hydrocarbon ring containing one or more double bonds in ring.For example, C3-C6 cycloalkenyl groups refer to there is 3 To the cycloalkenyl group of 6 member carbon atoms.In some embodiments, cycloalkenyl group has a carbon-to-carbon double bond in ring.Other In embodiment, cycloalkenyl group has more than one carbon-to-carbon double bond in ring.Cyclenes basic ring is not fragrant.Cycloalkenyl group is single Member ring systems.Cycloalkenyl group optionally can be replaced by one or more substituents as defined herein.Cycloalkenyl group includes cyclopropanyl, ring Cyclobutenyl, cyclopentenyl, cyclohexenyl group and cyclohexadienyl.

" aryl " refers to aromatic hydrocarbon ring system.Aryl is single ring systems or bicyclic system.Monocyclic aromatic rings refer to phenyl.It is bicyclic Aromatic ring refer to naphthyl and wherein phenyl with 5, the ring of the cycloalkyl of 6 or 7 member carbon atoms or the fusion of cyclenes basic ring.Aryl can Optionally to be replaced by one or more substituents as defined herein.

Term " heteroaryl " or " aromatic heterocycle " include substituted or unsubstituted aromatic monocyclic structure, preferably 5 to 7- Yuan of rings, more preferably 5 to 6 yuan of rings, its ring structures include at least one hetero atom, preferably 1-4 hetero atom, more preferably 1 or 2 hetero atom.Term " heteroaryl " also includes the member ring systems with one or two ring, and wherein at least one ring is heteroaryl Fragrant ring (such as) and another ring can be cycloalkyl, cycloalkenyl group, cycloalkynyl radical, aromatic carbocyclic, heteroaryl and/or heterocyclic radical.Heteroaryl Including for example, pyrroles, furans, thiophene, imidazoles, oxazoles, thiazole, pyrazoles, pyridine, pyrazine, pyridazine and pyrimidine.

Term " heterocycle " used herein and " heterocycle " refer to non-aromatic or aromatic rings, and it is comprising one or more Hetero atom selected from such as N, O, B and S atom, preferably N, O or S.Term " heterocycle " includes " aromatic heterocycle " and " non-aromatic miscellaneous Both rings ".Heterocycle includes 4-7 unit monocycles and 8-12 membered bicyclics.Heterocycle include bicyclic molecule, one of them, two or three or Multiple atoms are shared by two rings.Each ring of bicyclic heterocycle can be selected from non-aromatic ring and aromatic rings." fusion is miscellaneous for term Ring " refers to bicyclic heterocycle, wherein each ring shares two adjacent atoms with other rings.Each ring of annelated heterocycles may be selected from Non-aromatic ring and aromatic rings.In exemplary embodiment, aromatic rings (such as pyridine radicals) can be fused to non-aromatic ring or Aromatic rings, such as hexamethylene, pentamethylene, pyrrolidines, DHF or cyclohexene." heterocycle " group includes, for example, piperazine Pyridine, piperazine, pyrrolidines, morpholine, pyrimidine, benzofuran, indoles, quinoline, lactone and lactams.Exemplary " annelated heterocycles " bag Include benzodiazepineIndoles, quinoline, purine and 4,5,6,7- tetrahydro benzos [d] thiazole." heterocycle " can be at any one or more The position can with hydrogen atom is substituted.

It is " monocyclic " to include 5-7 members aromatic carbocyclic or heteroaryl, 3-7 members cycloalkyl or cycloalkenyl group, and 5-7 member nonaromatic heterocycles Base.Exemplary monocyclic groups include substituted or unsubstituted heterocycle or carbocyclic ring, for example, thiazolyl, oxazolyl, oxazinyls, Thiazinyl, dithian base, alkyl dioxin, isoxazolyls, isothiazolyl, triazolyl, furyl, tetrahydrofuran base, two Hydrogen furyl, pyranose, tetrazole radical, pyrazolyl, pyrazinyl, pyridazinyl, imidazole radicals, pyridine radicals, pyrrole radicals, pyrrolin base, Pyrrolidinyl, piperidyl, piperazinyl, pyrimidine radicals, morpholinyl, tetrahydro-thienyl, thienyl, cyclohexyl, cyclopenta, cyclopropyl, Cyclobutyl, suberyl, azetidinyl, oxetanyl, thiirane base, oxirane base, aziridine Base and thiomorpholine base.

" member atoms " refer to the one or more atoms to form chain or ring.When in chain and there is more than one member in ring Atomic time, each member atoms member atoms covalent bond adjacent with chain or ring.Constitute the original of chain or the substituent on ring Son is not the member atoms in chain or ring.

It is " optionally substituted " expression group, such as alkyl, alkenyl, alkynyl, aryl, cycloalkyl, cycloalkenyl group, Heterocyclylalkyl or Heteroaryl can be unsubstituted, or the group can replace one or more substituents as herein defined.

As used herein, " substituted " refers to the hydrogen atom in atom in addition to hydrogen or molecule substitution structure.Refer to " substitution " during group represents that the one or more hydrogen atoms being connected with the member atoms in group are selected from defined substitution The substituent of base group is substituted.Commutable atom such as " commutable nitrogen " is the original that hydrogen atom is carried with least one resonance form Son.Hydrogen atom can be by other atom or substituent group, such as CH₃Or OH groups.For example, if nitrogen is combined with hydrogen atom, Then the nitrogen in piperidines molecule is commutable.For example, if the nitrogen of piperidines is bonded to atom in addition to hydrogen, nitrogen is inadvisable Generation.It can not replaced with atom of any resonance form with hydrogen atom.It should be appreciated that term " substituted " includes Such Implicit Conditions：This substitution meets the chemical valence of the permission of the atom replaced and substituent, and replaces generation steady Fixed compound (it will not be reset spontaneously for example by hydrolysis, cyclisation or eliminate the compound that converted, and enough It is solid to be separated with being resistant to from reactant mixture).When pointing out that group can contain one or more substituents, one in group Individual or multiple (optionally) member atoms can be substituted.As long as in addition, this substitution meets the chemical valence of the permission of atom, then Single member atom in the group can be replaced by more than one substituent.For each substitution or optionally substituted base Group, suitable substituent is defined herein.

The present invention relates to the combination of substituent and variable, condition is to form stable compound.Term as used herein is " steady Fixed " refer to that compound has its preparation of permission and the integrality of the compound can be made to maintain enough time for herein The sufficiently stable property of purpose be described in detail.

Deuterated compound

Compound disclosed herein is also included through part and completely deuterated variant.In some embodiments, deuterium The variant in generation can be used for dynamics research.The site that the D-atom is present may be selected in those of ordinary skill in the art.

Present invention additionally comprises formula (I) compound or the various deuterated forms of its pharmaceutically acceptable salt.Connect with carbon atom The each available hydrogen atom connect can be replaced independently by D-atom.Those of ordinary skill in the art will know how synthesis originally Invention formula (I) to (II) compound deuterated form.For example, deuterated material such as alkyl can be prepared by routine techniques (see, for example,：Derived from Aldrich Chemical Co., Milwaukee, WI, catalog number (Cat.No.) 489,689-2 methyl-d3- amine).

Isotope

Present invention additionally comprises isotope marks and identical compound described in formula (I) and (II), but it is the fact that It is different：One or more atoms are different from most common atomic mass or mass number in nature by atomic mass or mass number Atom is substituted.May be incorporated into the example of the isotope of the compounds of this invention includes hydrogen, carbon, nitrogen, oxygen, fluorine, the isotope of iodine and chlorine, example Such as³H、¹¹C、¹⁴C、¹⁸F、¹²³I or¹²⁵I。

The medicine of the compounds of this invention of other isotopes containing above-mentioned isotope and/or other atoms and the compound Acceptable salt is within the scope of the invention on.The compound of the isotope marks of the present invention, for example, mixed radioactivity Isotope is such as³H or¹⁴C those compounds can be used for medicine and/or substrate tissue measure of spread.Due to easily prepared and can examine The property surveyed, it is tritiated i.e.³H and carbon-14 are¹⁴C isotopes are particularly preferred.¹¹C and¹⁸F isotopes in PET, (break by positron emission Layer photography) in it is particularly useful.

Purity

Because the compound of the present invention is intended to be used in pharmaceutical composition, it is readily appreciated that, each of which is preferably with base Pure form is provided in sheet, and for example, at least 60% is pure, and more suitably at least 75% is pure, and preferably at least 85%, especially at least 98% pure (% is by weight).The not pure preparation of compound can be used for preparing the purer form used in pharmaceutical composition.

Salt

In certain embodiments, the compound of Formulas I or its pharmaceutically acceptable salt can contain acidic functionality.At certain In other a little embodiments, the compound of Formulas I can contain basic functionality.Thus, it will be understood by those skilled in the art that can prepare The salt of compound of formula I.In fact, in certain embodiments of the invention, the salt of compound of formula I is relative to respective free alkali Or free acid be probably preferably as, such as such salt can assign the molecule bigger stability or dissolubility, thus Be conducive to being formulated as formulation.

Due to its potential use in medicine, the salt suitably from pharmaceutically acceptable salt of formula (I) compound.Properly Pharmaceutically acceptable salt include Berge, Bighley and Monkhouse, J.Pharm.Sci. (1977) 66, the 1-19 pages Described in those.

The present invention also includes the salt of compound as described herein, particularly pharmaceutically acceptable salt.With it is enough it is acid, Alkalescence or acid and both functional groups of alkalescence the compounds of this invention enough, can be with many inorganic bases and inorganic with having Any of machine acid reacts forming salt.Or, intrinsic electrically charged compound (such as with season nitrogen compound) can be with fitting When counter ion counterionsl gegenions (for example, halogen ion such as bromide ion, chlorion or fluorine ion, particularly bromide ion) forming salt.

The acid for being typically used to form acid-addition salts is inorganic acid, such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid Deng, and organic acid, such as p- toluenesulfonic acid, methanesulfonic acid, oxalic acid, p- bromophenyl-sulfonic acid, carbonic acid, butanedioic acid, citric acid, Benzoic acid, acetic acid etc..The example of such salt includes sulfate, pyrosulfate, bisulfate, sulphite, acidic sulfurous acid Salt, phosphate, mono-hydrogenphosphate, dihydric phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetic acid Salt, propionate, caprate, caprylate, acrylates, formates, isobutyrate, caproate, enanthate, propiolate, oxalic acid Salt, malonate, succinate, suberate, sebacate, fumarate, maleate, butine -1,4- diacid salts, oneself Alkynes -1,6- diacid salts, benzoate, chloro benzoate, methyl benzoic acid salt, dinitro-benzoate, hydroxy benzoate, first P-methoxybenzoic acid salt, phthalate, sulfonate, xylenesulfonate, phenyl acetate salt, phenylpropionic acid salt, PB, lemon Hydrochlorate, lactate, gamma hydroxybutyrate, glycollate, tartrate, mesylate, propane sulfonic acid salt, naphthalene -1- sulfonate, naphthalene - 2- sulfonate, mandelate etc..

Base addition salts include being derived from inorganic base, such as ammonium or alkali metal or alkaline earth metal hydroxide, carbonate, carbonic acid Those salt of hydrogen salt etc..Therefore such alkali that can be used for preparing salt of the present invention including sodium hydroxide, potassium hydroxide, ammonium hydroxide, Potassium carbonate etc..

" enantiomeric excess " or " ee " is a kind of enantiomter relative to another excess, as a percentage.Therefore, When two kinds of enantiomters are present in racemic mixture with equal amount, the enantiomeric excess is zero (0%ee).But It is, if a kind of enrichment of enantiomter is so that when constituting the 95% of the product, then the enantiomeric excess is 90%ee (rich The amount of the enantiomter of collection, 95%, subtract the amount of another enantiomter, 5%).

" enantiomer enrichment " refers to product of its enantiomter excess of greater than zero.For example, enantiomter enrichment refers to Its enantiomeric excess is more than 50%ee, the product more than 75%ee or more than 90%ee.

" enantiomer-pure " refers to that its enantiomeric excess is 99%ee or bigger product.

" pharmaceutically acceptable " refers in rational medical judgment scope, it is adaptable to contacted with the tissue of humans and animals Without excessive toxicity, excitant or other problemses or complication, and those chemical combination with rational interests/Hazard ratio Thing, material, composition and formulation.

Formula (I) compound or its pharmaceutically acceptable salt can be (also referred to as chiral containing one or more asymmetric centers Center), therefore individually enantiomer, the form of diastereomer or other stereoisomers or can be deposited in the form of its mixture .

Chiral centre such as asymmetric carbon atom also is present in substituent such as alkyl.When being present in Formulas I or this paper institutes When the spatial chemistry of chiral centre in any chemical constitution illustrated is not indicated, the structure is intended to include all single solids Isomers and its all mixtures.

Therefore, formula (I) compound or its pharmaceutically acceptable salt containing one or more chiral centres may be used as Racemic mixture, non-enantiomer mixture, enantiomer enrichment mixture, diastereomer enrichment mixture or as mapping Body and the pure single stereoisomer of diastereomer.

The compound of (I) containing one or more asymmetric centers or the independent solid of its pharmaceutically acceptable salt are different Structure body can be split by method known to those skilled in the art.For example, the fractionation can be carried out as follows：(1) by being formed Diastereomeric salt, compound or other derivatives；(2) by the selective reaction with stereoisomer specific reagent, For example pass through enzymatic oxidation or reduction；Or (3) pass through the gas liquid chromatography or liquid chromatogram in chiral environment, the chiral ring Border is for example in chiral support (such as the silica gel for being connected with chiral ligand) or in the presence of chiral solvent.Those skilled in the art will It will be appreciated that when required stereoisomer is changed into another diastereomeric salt, it is necessary to which other steps discharge required shape Formula.Or, specific stereoisomer can by using optical activity reagent, matrix, catalyst or solvent asymmetric syntheses Method is synthesized, or a kind of enantiomter is changed into another enantiomter by asymmetric transformation.

When disclosed compound or its salt is named or described with structure, it will be appreciated that compound or its salt includes Its solvate (particularly its hydrate) can in crystalline form, non-crystalline forms or its mixture be present.Compound or salt Or its solvate (particularly hydrate) can also show polymorphism (ability existed with different crystal forms). These different crystal forms are commonly known as " polymorph ".

In consideration of it, (that is, it includes its different polymorphs, anhydrous form, solvate or water to the salt form of the present invention Compound) expression characteristicses polymorphism.As this area routinely understand as, polymorphism be defined as compound with More than one different crystallization or the ability of " polymorphic " species crystallization.Polymorph is defined as the solid crystal of compound Phase, its at least two different arrangement with the compound molecule of solid-state or polymorphic forms.

The polycrystalline form of any given compound (compound for including the present invention) is defined by identical chemical formula or constitute, And it is two kinds of different compounds for the chemical constitution as crystalline texture.The compound may be in respective lattice Accumulation, different in terms of geometry arrangement.

It should be appreciated that when being named by structure or being described, disclosed compound or solvate (particularly its hydration Thing) also include its all polymorph.Polymorph has an identical chemical composition, but accumulation in crystalline solid state, geometry row Row are different with terms of other descriptive characteristics.

In view of above-mentioned, chemistry and/or physical property or characteristic change with each different polycrystalline form, and it may include molten Xie Du, fusing point, density, hardness, crystal shape, electrical and optical properties, steam pressure, the change of stability etc..

When solvent molecule is incorporated into the lattice structure of compound molecule in crystallization process, this hair can also be formed The solvate and/or hydrate of bright crystalline salt form.For example, the solvate forms of the present invention can introduce following article institute The nonaqueous solvents stated such as methanol etc..Hydrate forms are that water is mixed to the solvate form thereof in lattice as solvent.

The anhydrous crystal structure referred to without the recrystallisation solvent repeated in lattice on solid multi-crystalline type phenomenon.So And, crystalline material can be reversible adsorption that is porous and can showing water outlet.

Term and definition

Part 2

1. definition

Following term and phrase for this paper should have in the implication hereafter described.Unless otherwise defined, Suo Youyu Technology used herein has the identical meanings being generally understood that with those of ordinary skill in the art with scientific terminology.

The terms " medicament (agent) " are used to represent compound, the mixture of compound, large biological molecule (for example Nucleic acid, antibody, albumen or part thereof such as peptide) or from biomaterial such as bacterium, plant, (the particularly lactation of fungi or animal Animal) extract made from cell or tissue.

Term " bioavailable " is well known in the art when being related to compound, and considered be related to chemical combination Thing form, wherein to drug compound amount all or part of subject that can be administered or patient absorb, introduce Or be otherwise utilized in a physiologically.

" part of the biological activity of Sirtuin " refers to bioactivity (such as deacetylated ability) A part for the Sirtuin of (" catalytic activity ").The catalytical active part of Sirtuin can adjust egg comprising silence White Core domain.Numbering NP_036370 SIRT1 catalytical active part, including NAD are logged in GenBank⁺With reference to Domain and substrate-binding domain, for example, may include but be not limited to the amino acid that GenBank logs in numbering NP_036370 240-664 or 240-505, it is as coded by GenBank login numberings NM_012238 polynucleotides.Therefore, this region is sometimes Referred to as Core domain.SIRT1 other catalytical active parts (being also sometimes referred to as Core domain) include GenBank Numbering NP_036370 substantially amino acid 261 to 447 is logged in, it is logged in numbering NM_012238 nucleotides 834 by GenBank Coded by 1394；GenBank logs in numbering NP_036370 substantially amino acid 242 to 493, and it is logged in by GenBank and numbered Coded by NM_012238 nucleotides 777 to 1532；Or GenBank log in numbering NP_036370 substantially amino acid 254 to 495, it is as coded by GenBank login numberings NM_012238 nucleotides 813 to 1538.SIRT1 another " biology Activity " partly logs in numbering NP_036370 amino acid 62-293 or 183-225 for GenBank, and it is included for compound knot Close the N- terminal domains of the important Core domain in site.

Term " companion animals " refers to cat and dog.Term " dog " for this paper refers to any member of Canidae family, its In have many different cultivars.Term " cat " refers to cats, including raises and train other members of cat and cat family Felis.

" diabetes " refer to hyperglycaemia or ketoacidosis, and because of long term hyperglycemia state or glucose tolerance attenuating institute Caused chronic, general metabolic disorder." diabetes " comprising the disease I and II type (adult-onset diabetes or NIDDM) two kinds of forms.The hazards of diabetes include following factors：Man's waistline is more than 40 inches or woman's waistline exceedes 35 inches, blood pressure is 130/85mmHg or more, and triglyceride is higher than 150mg/dl, and fasting blood-glucose is more than 100mg/dl, or Man's HDL is less than 40mg/dl or woman's HDL is less than 50mg/dl.

Term " ED₅₀" be effective dose well known in the art measurement.In some embodiments, ED₅₀Refer to that medicine is produced Its raw peak response or effect 50% when dosage, or in 50% test subject or product (such as tissue of separation Or cell) the middle dosage for producing predetermined response.Term " LD₅₀" be lethal dose well known in the art measurement.In some implementations In mode, LD₅₀Refer to that medicine makes the 50% lethal dosage of test subject.Term " therapeutic index " is well known in the art Term, refers to the therapeutic index of medicine, is defined as LD₅₀/ED₅₀。

Term " hyperinsulinemia " refers to state of the insulin level higher than normal value in individual blood.

Term " insulin resistance " refers to a kind of such state, wherein relative to without the tested of insulin resistance For biological response in person, normal amount insulin produces the state less than normal (subnormal) biological response.

" insulin resistance symptoms " discussed in this article refer to as caused by insulin resistance or any disease for facilitating or Illness.Example includes：Diabetes, obesity, metabolic syndrome, insulin resistance syndrome, syndrome X, insulin resistance, height Blood pressure, hyperpiesia, high blood cholesterol, dyslipidemia, hyperlipidemia, atherosclerosis disease, including apoplexy, coronary artery The former mass formed by blood stasis of disease or miocardial infarction, hyperglycemia, hyperinsulinemia and/or hyperinsulinism, glucose tolerance are bad, delay Insulin releasing, diabetic complication, including coronary heart diseases and angina pectoris, congestive heart failure, apoplexy, dull-witted cognitive function, Retinopathy, peripheral nerve disease, nephrosis, glomerulonephritis, glomerulosclerosis, nephrotic syndrome, hypertensive nephrosclerosis, one A little types of cancer (such as carcinoma of endometrium, breast cancer, prostate cancer and colon cancer), complications of pregnancy, female reproductive health are bad (such as irregular menstruation, irregularly infertility, ovulation, polycystic ovary syndrome (PCOS)), lipodystrophia, cholesterol are related Illness such as gall stone, cholecystitis and cholelithiasis, gout, obstructive sleep apnea and breathing problem, osteoarthritis, and Bone loss, e.g. particularly osteoporosis.

Term " livestock animals " refers to the quadruped through domestication, and it includes those and raised for meat with all kinds of byproducts Animal, such as bovine includes other members of ox and Bos, porcine animals, including raise and train that pig and pig belong to it is other into Member, sheep class animal, including sheep belong to other members of (genus Ovis) with sheep, raise and train other members of goat and Capra； For example be used as the quadruped through domestication that heavy burden beast is raised, such as horse class animal for particular task, including raise and train horse with Other members of equine Equus.

Term " mammal " is known in the art, and Exemplary mammals include people, primate, livestock animals (including ox class, pig class etc.), companion animals (such as canine, cat class) and rodent (such as mouse and rat).

" obesity " individual or the individual with obesity, the body mass index (BMI) for referring generally to have is at least 25 or more Individual.Obesity can be related with or without insulin resistance.

Term " through parenteral " and " being administered through parenteral " be it is art-recognized, and duodenum 12 administration with it is local Mode of administration beyond administration, typically via injection, and including but not limited to：Intravenous, intramuscular, intra-arterial, intrathecal, capsule Interior, socket of the eye is interior, intracardiac, intradermal, intraperitoneal, transtracheal, under subcutaneous, epidermis, under intra-articular, capsule, under arachnoid, in backbone and chest Intraosseous injection and administered by infusion.

" patient ", " subject ", " individual " or " host " refers to people or non-human animal.

Term " pharmaceutically acceptable carrier " is art-recognized, and refers to and carry or transport any body combination Thing or its component relevant pharmaceutically acceptable material, composition or medium, such as liquid or solid filler, dilution Agent, excipient, solvent or encapsulating material.Various carriers with regard to its can be compatible with main component or its component meaning for it is necessary For " acceptable ", and it is harmless to patient.Can be used as some examples of the material of pharmaceutically acceptable carrier includes：(1) Carbohydrate, such as lactose, dextrose and saccharose；(2) starch, such as cornstarch and farina；(3) cellulose derives with it Thing, such as sodium carboxymethylcellulose, ethyl cellulose and cellulose acetate；(4) powdered tragcanth；(5) malt；(6) it is bright Glue；(7) talcum powder；(8) excipient, such as cocoa butter and suppository wax；(9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame Sesame oil, olive oil, corn oil and soybean oil；(10) glycols, such as propane diols；(11) polyalcohols, such as glycerine, sorbose Alcohol, mannitol and polyethylene glycol；(12) esters, such as ethyl oleate and ethyl laurate；(13) agar；(14) buffer, Such as magnesium hydroxide and aluminium hydroxide；(15) alginic acid；(16) without heat source water；(17) isotonic saline solution；(18) Ringer's solution； (19) ethanol；(20) phosphate buffer solution；And (21) other non-toxic compatible materials for pharmaceutical preparation.

Term " prevention " is well known in the art, and is being related to illness (such as local recurrence (such as pain)), disease (example Such as cancer), syndrome (such as heart failure) or any other medical conditions in use, it is well known in the present art, bag Administration composition is included, compared with not receiving the subject of said composition, administration composition reduces medical science disease in subject The frequency of the symptom of disease delays its breaking-out.Therefore, the prevention of cancer includes, for example, receiving preventative-therapeutic patient The quantity (relative to untreated control crowd) of detectable cancer growth is reduced in group, and/or in the crowd for the treatment of The appearance (compared with untreated control crowd) of detectable cancer growth is delayed, for example, with statistics and/or clinically significant Amount.The prevention of infection includes, for example, the quantity that the infection of diagnosis is reduced in the crowd for the treatment of (compares people with untreated Faciation ratio), and/or delay in the crowd for the treatment of the breaking-out of infection symptoms (compared with untreated control crowd).Pain Prevention include, for example, reducing the degree for the pain being aware of by the subject in the crowd for the treatment of or delaying The sensation (compared with untreated control crowd).

Term " preventative " or " therapeutic " treatment are well known in the art, and refer to drug administration to host.If Undesirable illness (for example, host animal disease or other be not intended to state) there is clinic to show before be administered, then The treatment is preventative, i.e., its protection host will not develop into this and be not intended to illness, and if be not intended to after illness shows It is administered, then the treatment is curative (is intended to reduce, improve or maintain existing to be not intended to illness or be generated by it Side effect).

Term " no thermal source (pyrogen-free) " refers to that the group will not be administered in thermal source content when being related to composition Cause illeffects (for example, stimulation, heating, inflammation, diarrhoea, respiratory distress, endotoxic shock etc.) in the subject of compound Composition.For example, term meaning includes the endotoxic composition free or substantially free of such as lipopolysaccharides (LPS).

" the replicating the life-span " of cell refers to as the daughter cell quantity produced by single " mother cell ".On the other hand, " sequential declines Always (chronolo gical aging) " or " sequential life-span " refers to that the nondividing cell of a group is still kept when being removed nutrient The time span of survival." increase cell survival " or " extension cell survival " refers to increase when applied to cell or organism As the daughter cell quantity produced by a cell；Increase cell or organism to resisting stress and to antibody Monoclonal (such as to DNA, egg It is white stress and damage) ability；And/or increase cell or organism for example stress (such as heat shocks, osmotic pressure in special conditions of contract Power, high energy radiation, chemical induction stress, DNA damage, insufficient salt level, insufficient nitrogen level or insufficient Nutrient level) under the state of survival and existence there is the ability of long period.Using method specifically described herein, the life-span can increase Plus at least about 10%, 20%, 30%, 40%, 50%, 60%, 20% to 70%, 30% to 60%, 40% to 60% or with On.

" compound of regulation Sirtuin " refers to increase Sirtuin level, and/or increase silence regulation At least one active compound of albumen.In an exemplary embodiment, the compound of regulation Sirtuin can make At least one bioactivity increase at least about 10%, 25%, 50%, 75%, 100% or more of Sirtuin.Silence The illustrative bioactivity of regulatory protein includes the deacetylated of such as histone and p53；Extend the life-span；Increase genome Stability；Silence transcription；And the separation of regulation and control mother cell and the oxidized albumen of careful intercellular.

Albumen is including deacetylated, for example, acetylated peptide substrate is deacetylated.

" Sirtuin " refers to a member of the deacetylation zymoprotein family of Sirtuin or preferably refers to sir2 A member of family, it includes yeast Sir2 (GenBank logs in numbering P53685), Caenorhabditis elegans Sir-2.1 (GenBank Log in numbering NP_501912) and people SIRT1 (GenBank logs in numbering NM_012238 and NP_036370 (or AF083106)) With SIRT2 (GenBank logs in numbering NM_012237, NM_030593, NP_036369, NP_085096 and AF083107) egg In vain.Other family members include be referred to as " HST genes " (Sir2 homologue) four kinds of other yeast Sir2 sample genes be HST1, HST2, HST3 and HST4, and five kinds of other human homologues hSIRT3, hSIRT4, hSIRT5, hSIRT6 and hSIRT7 (Brachmann et al., (1995) Genes Dev.9：2888 and Frye et al. (1999) BBRC 260：273).

" SIRT1 albumen " refers to a member of the Sir2 families of the deacetylase of Sirtuin.In some embodiment party In formula, SIRT1 albumen includes yeast Sir2 (GenBank logs in numbering P53685), Caenorhabditis elegans Sir-2.1 (GenBank Log in numbering NP_501912), people SIRT1 (GenBank login numbering NM_012238 or NP_036370 (or AF083106)), Mouse SIRT1 (GenBank logs in numbering NM_019812 or NP_062786) and its equivalent and fragment.In another embodiment party In formula, SIRT1 albumen includes polypeptide, its include by or substantially by GenBank log in numbering NP_036370, NP_501912, The sequence that amino acid sequence shown in NP_085096, NP_036369 or P53685 is constituted.SIRT1 albumen is included under containing The all or part of polypeptide and its functional fragment of row amino acid sequence：GenBank logs in numbering NP_036370, NP_ 501912nd, the amino acid sequence shown in NP_085096, NP_036369 or P53685；GenBank logs in numbering NP_ 036370th, shown in NP_501912, NP_085096, NP_036369 or P53685 have 1 to about 2,3,5,7,10,15, 20th, the amino acid sequence of 30,50,75 or more conservative amino acids substitution；Numbering NP_036370, NP_ is logged in GenBank 501912nd, NP_085096, NP_036369 or P53685 at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98% Or 99% identical amino acid sequence and its functional fragment.Polypeptide in the present invention also includes GenBank and logs in numbering NP_ 036370th, NP_501912, NP_085096, NP_036369 or P53685 homologue (straight homologues (orthologs) with Lateral homologue (paralogs)), variant or fragment.

" SIRT2 albumen " used herein, " SIRT3 albumen ", " SIRT4 albumen ", " SIRT5 albumen ", " SIRT6 albumen " " SIRT7 albumen " refer to the Sirtuin of other mammals (such as people) deacetylation zymoprotein (its with SIRT1 albumen homologies), especially in about 275 conservative catalyst structure domains.For example, " SIRT3 albumen " refers to that silence is adjusted The member of the deacetylation zymoprotein family (itself and SIRT1 albumen homologies) of albumen.In one embodiment, SIRT3 albumen Including people SIRT3 (GenBank logs in numbering AAH01042, NP_036371 or NP_001017524) and mouse SIRT3 (GenBank logs in numbering NP_071878) albumen, and its equivalent and fragment.In some embodiments, SIRT4 albumen bag Include people SIRT4 (GenBank logs in numbering NM_012240 or NP_036372).In some embodiments, SIRT5 albumen includes People SIRT5 (GenBank logs in numbering NM_012241 or NP_036373).In some embodiments, SIRT6 albumen includes people SIRT6 (GenBank logs in numbering NM_016539 or NP_057623).In another embodiment, SIRT3 albumen includes many Peptide, it includes by or substantially logs in numbering AAH01042, NP_036371, NP_001017524 or NP_ by GenBank The sequence that amino acid sequence shown in 071878 is constituted.SIRT3 albumen include the whole containing following amino acid sequences or The polypeptide and its functional fragment of a part：GenBank log in numbering AAH01042, NP_036371, NP_001017524 or Amino acid sequence shown in NP_071878；GenBank log in numbering AAH01042, NP_036371, NP_001017524 or The ammonia replaced with 1 to about 2,3,5,7,10,15,20,30,50,75 or more conservative amino acids shown in NP_071878 Base acid sequence；With GenBank log in numbering AAH01042, NP_036371, NP_001017524 or NP_071878 at least 60%, 70%th, 80%, 90%, 95%, 96%, 97%, 98% or 99% identical amino acid sequence and its functional fragment.The present invention In polypeptide also include GenBank and log in the homologous of numbering AAH01042, NP_036371, NP_001017524 or NP_071878 Thing (straight homologues and lateral homologue), variant or fragment.In some embodiments, SIRT3 albumen is included with mitochondria The SIRT3 protein fragments that matrix processing peptidase (MPP) and/or mitochondria intermediate peptase (MIP) are cracked and produced.

Term " stereoisomer " used herein is well known in the art, and refers to two or more and have phase With molecular composition and only spatially in the different isomers of the three-dimensional arrangement of their atom any one.When being used in the text When describing compound or general formula compound, stereoisomer includes any portion or whole compound of the compound.For example, Diastereoisomer and enantiomter are stereoisomers.

Term " systemic applications " and " being capapie administered " are well known in the art, and refer to theme composition, treatment Agent or other materials enteral administration or parenteral.

Term " dynamic isomer " used herein is well known in the art, and refers to what is caused by tautomerism Any one in structure that may be present, it refers to stereo isomers form, and wherein structure can be arranged with two or more constructions Row form is present, for the position for being especially coupled to the hydrogen of oxygen.When in the text for describing compound or general formula compound When, further understanding " dynamic isomer " is easy mutually phase in version and is in poised state.For example, keto-acid and enol form change Isomers exists in certain proportion, and this depends on the equilbrium position for specified criteria or condition group：

Term " therapeutic agent " is well known in the art, and refer to can be in the life topically or systemically worked in subject Thing, physiologically or pharmacologically active material.The term, which is also referred to, to be intended to for diagnosing, curing, mitigate, treat or prevent disease Any material, or for promoting the desired body or Mental development and/or the arbitrary substance of situation of animal or people.

Term " therapeutic effect " is well known in the art, and refers to animal (particularly mammal, and more particularly People) in the beneficial topically or systemically effect that is produced by pharmacological active substance.Phrase " therapeutically effective amount " refers to, makes material Reasonable benefit/Hazard ratio that treatment can be suitable for produces the amount of the locally or systemically effect desired by certain.The treatment of material has Effect amount will have with being intended to treated subject with illness, the body weight of subject and age, the severity of illness, administering mode etc. Changed, it can be readily determined by those of ordinary skill in the art.For example, some compositions as described herein can be suitable for controlling Reasonable benefit/Hazard ratio for the treatment of is applied so as to produce the sufficient amount of desired effect to be administered.

" treatment " illness or disease refer to cure and improve the illness or at least one symptom of disease.

Term " vision impairment " refers to that eyesight weakens, and often only part is reversible or not when (such as operation) being treated for it It is reversible.The vision impairment of especially severe is referred to as " blind " or " visual loss ", and it refers to that eyesight completely loses, eyesight is worse than 20/200 So that can not improve via correcting lens, or the visual field is less than 20 degree of diameters (10 degree of radiuses).

Abbreviation and symbol

When describing the present invention, chemical element is differentiated according to the periodic table of elements.Abbreviation used herein and symbol symbol The technical staff of combination and field of biology is to these abbreviations and the conventional purposes of symbol.

Specifically, following abbreviation is used in the whole text in embodiment and specification：

G (gram) mg (milligram)；

Kg (kilogram) μ g (microgram)；

L (liter) mL (milliliter)；

μ L (microlitre) psi (pound/square inch)；

M (mole every liter) mM (mM every liter)；

μM (every liter of micromole) nM (every liter of nanomole)；

PM (every liter of picomole) nm (nanometer)；

Millimeter (millimeter) weight (weight)；

N (standard) CFU (CFU)；

I.V. (intravenous) Hz (hertz)；

MHz (megahertz) moles (mole)；

Mmol (mM) RT (room temperature)；

Min (minute) h (hour)；

B.p. (boiling point) TLC (thin-layer chromatography)；

Tr (retention time) RP (anti-phase)；

MeOH (methanol) i-PrOH (isopropanol)；

TEA (triethylamine) TFA (trifluoroacetic acid)；

TFAA (TFAA) THF (tetrahydrofuran)；

DMSO (dimethyl sulfoxide (DMSO)) EtOAc (ethyl acetate)；

DME (1,2- dimethoxy-ethanes) DCM (dichloromethane)；

DCE (dichloroethanes) DMF (N,N-dimethylformamide)；

DMPU (N, N'- dimethyl propylene alkenyl urea) CDI (1,1- carbonyl dimidazoles)；

IBCF (isobutyl chlorocarbonate) AcOH (acetic acid)；

HOAt (1- hydroxyl -7- azepines BTA)；

THP (oxinane) NMM (N-methylmorpholine)；

Pd/C (palladium/carbon) MTBE (t-butyl methyl ether)；

HOBT (I-hydroxybenzotriazole) mCPBA (metachloroperbenzoic acid)；

EDC (1- [3- dimethylaminos] propyl group] -3- ethyl-carbodiimide hydrochlorides)；

Boc (tertbutyloxycarbonyl) FMOC (9- fluorenylmethoxycarbonyl groups)；

DCC (dicyclohexylcarbodiimide) CBZ (benzyloxycarbonyl group)；

Ac (acetyl group) atm (atmospheric pressure)；

TMSE (2- (trimethyl silyl) ethyl) TMS (trimethyl silyl)；

TIPS (triisopropylsilyl) TBS (t-butyldimethylsilyl)；

DMAP (4-dimethylaminopyridine) BSA (bovine serum albumin(BSA))；

NAD (NADH)；

HPLC (high pressure liquid chromatography)；

LC/MS (liquid chromatography/mass spectrometry)；

BOP (double (2- oxo -3- oxazoles alkyl) secondary phosphonic chloride)；

TBAF (tetra-n-butyl ammonium fluoride)；

HBTU (O- BTA -1- bases-N, N, N ', N '-tetramethylurea hexafluorophosphate)；

HEPES (4- (2- ethoxys) -1- piperazine ethanesulfonic acids)；

DPPA (diphenyl phosphoryl azide) LAH (lithium aluminium hydride reduction)；

FHNO3 (smoke HNO₃) NaOMe (sodium methoxide)；

EDTA (ethylenediamine tetra-acetic acid)；

TMEDA (N, N, N', N'- tetramethyl -1,2- ethylenediamines)；

NBS (N- bromines succinimide) DIPEA (diisopropylethylamine)；

Dppf (1,1'- bis- (diphenylphosphino) ferrocene)；And

NIS (N-iodosuccinimide).

All ethers referred to are all ether, and salt solution refers to NaCl saturated aqueous solution.

Synthetic schemes and it is typically prepared method

The present invention also relates separately to prepare formula (I) to (IV) compound, corresponding analog (that is, in R²Position has hydrogen Substituted analog) and/or its midbody compound method.

Formula (I) to (IV) compound, corresponding analog is (that is, in R²Position has the analog that hydrogen replaces), and/or its Midbody compound or its pharmaceutically acceptable salt, by using the synthetic operation shown in following scheme or can be utilized ripe The knowledge of experienced organic chemist is obtained.

The synthesis provided in these schemes (I) to (VI) is applied to of the invention change of the production with various different functional groups Compound, using suitable precursor, if necessary to suitably protecting, to realize the compatibility of the reaction with summarizing herein.Needed with backsight It is deprotected that there is provided the compound with substantially disclosed property.Although scheme is shown for compound, they are also solved Release the method illustrated for preparing the compounds of this invention.

Intermediate (being used for the compound for preparing the compounds of this invention) can also salt form presence.Accordingly, with respect to intermediate, Phrase " compound of formula (so-and-so numeral) " refers to compound or its pharmaceutically acceptable salt with the structural formula.

The present invention also relates separately to prepare formula (I) to (IV) compound, and corresponding analog is (that is, in R²Position has hydrogen Substituted analog), and/or its midbody compound, or its pharmaceutically acceptable salt method.

The compound of formula (I) to (II) is prepared using conventional organic synthesis respectively.The invention further relates to prepare formula (I) extremely (IV) compound, corresponding analog is (that is, in R²Position has the analog that hydrogen replaces), and/or its midbody compound, or The method of its pharmaceutically acceptable salt.

The compound of the present invention can be organised by using the synthetic operation shown in following scheme or using skilled The knowledge of scholar is obtained.

Suitable synthetic route is described in following General reactions scheme.

It is prepared by compound

According to another embodiment, the invention provides the method for preparing compound defined above.Compound can be with Synthesized using routine techniques.Advantageously, Material synthesis of these compounds generally by being readily available.

It is known in the art available for the synthesis chemical conversion and methodology for synthesizing compound described herein, and including Such as described in documents below those：R.Larock,Comprehensive Organic Transformations(1989)； T.W.Greene and P.G.M.Wuts,Protective Groups in Organic Synthesis,2d.Ed. (1991)；L.Fieser and M.Fieser,Fieser and Fieser's Reagents for Organic Synthesis(1994)；And L.Paquette, ed., Encyclopedia of Reagents for Organic Synthesis(1995)。

General operation

Scheme I

Reagent:(a)THF,NaHCO₃,45℃；(b)Fe,i-PrOH,HOAc,70℃；(c)LiAlH₄,THF,60℃；(d) POCl₃。

In aprotic solvent (such as THF, DMF , dioxanes), in the presence of alkali (to remove HCl), by commercially available chloro pyrrole There is provided regioselectivity addition compound product (I-3) with nucleophilicity amine (I-2) reaction for pyridine (I-1).Bronstead acid (HCl, HOAc) and in the presence of proton solvent, using the nitro functions of Fe (0) reducing substances (I-3) (referring to Bechamp reduction,Org React.2,428,1944).Other metals can be used, such as Sn realizes the reduction.It is formed in situ Intermediate amine substance at elevated temperatures with ester functional group reactionses formation cyclic amides I-4.Make strong hydride reducer Such as LiAlH₄Reacted with compound I-4, cause ester being reduced into corresponding alcohol, while by lactam reduction circlewise amine.It is this The reduction of type is well-known to those skilled in the art, sees H.C.Brown and S.Krishnamurthy, Tetrahedron, Tetrahedron,1979,35,567.Alcohol (I-5) and activated group (such as POCl₃) reaction can be formed and be readily obtained (facile) there is provided bicyclic amine compound (I-6) for leaving group.

Scheme II

Reagent:(a)Pd₂(dba)₃,X-Phos,K₃PO₄,ArB(OH)₂, dioxane/H₂O；(b)NBS,CHCl₃,60℃； (c)NCS,CHCl₃,60℃；(d)NIS,CHCl₃,60℃；(e) double (tetrafluoro boric acids of the fluoro- N'- chloromethyls triethylenediamines of N- Salt), trifluoromethanesulfonic acid, 60 DEG C.

Compound I-6 chlorine functional group and boric acid coupling are obtained into II-1 using Suzuki conjugation chemistries.Usually using all Such as Pd (PPh₃)₄Palladium (0) catalyst and inorganic basis such as K₂CO₃、Na₂CO₃Or K₃PO₄Containing ether solvents such as DME , dioxanes Or Suzuki sample couplings are carried out in THF aqueous mixture.The method of palladium mediated coupling is described in canonical reference handbook, for example " Organometallics in Synthesis " (are published) Schlosser by Wiley and sons.By compound II-1 with The fluoro- N'- chloromethyls triethylenediamines of electrophilic halogenation reagent such as NCS, NBS, NIS or N- double (tetrafluoroborates) are suitable Reacted in solvent, corresponding halogenated substances II-2 is obtained with regioselective manner.There are many methods to realize the halogen of aromatic ring Change, and be well-known to those skilled in the art.

Scheme III

Reagent:(a)Zn(CN)₂,Zn(OAc)₂,DPPF,Pd₂(dba)₃,DMF,110℃(b)CuCl,KOt-Bu,CF₃Si (CH₃)₃, 1,10- phenanthroline, DMPU, 35 DEG C of (c) CH₃B(OH)₂,K₃PO₄,X-Phos,Pd₂(dba)₃, dioxane/H₂O,75 ℃。

The intermediate that compound III-1 halogen group can use metal to mediate is replaced with various functional groups, is changed Compound III-2.Organic metal coupling is usually using palladium (0) catalyst such as Pd (PPh₃)₄Or other metals such as Cu or Sn, with alkali example Such as KOt-Bu or K₃PO₄Carried out in the mixture containing polar solvent such as DMPU, dioxanes or DMF.The method of palladium mediated coupling Described in canonical reference handbook, " Organometallics in Synthesis " are (by Wiley and by such as Schlosser Sons is published).The substitution for the metal catalytic that there are many methods to realize the halide on aromatic ring, and be art technology Known to personnel.

Scheme IV

Reagent:(a) TEA, triphosgene, DCM；(b) aniline, TEA, DCM；(c)DPPA,ArCO₂H,60℃；(d)Pd₂ (dba)₃,X-Phos,K₃PO₄, dioxane/H₂O。

By amine (IV-1) and acylating reagent such as triphosgene or carbonyl dimidazoles in aprotic solvent (DCM, CHCl₃, THF etc.) Middle reaction, obtains reactive acyl intermediate material (IV-2).In the presence of tertiary alkyl amine base, with aniline compound or alkylamine In-situ treatment reactivity acyl compounds (IV-2), form urea material (IV-3).Or, suitable carboxylic can be handled with DPPA Acid, it is then in situ at elevated temperatures to add amine (IV-1), produce urea (IV-3).This reaction is related to Curtius rearrangements (Org.React.3,3371946), and be well-known to those skilled in the art.Using Suzuki conjugation chemistries by urea IV-3 Chlorine functional group and boric acid be selectively coupled, obtain (IV-4).Usually using such as Pd (PPh₃)₄Palladium (0) catalyst with it is inorganic Alkali such as K₂CO₃、Na₂CO₃Or K₃PO₄Suzuki is carried out in the aqueous mixture containing ether solvents such as DME, dioxanes or THF Sample is coupled.The method of palladium mediated coupling is described in canonical reference handbook, such as Schlosser " Organometallics in Synthesis " (is published) by Wiley and sons.The reaction sequence can be overturned so that IV-1 is first in Suzuki conditions Lower reaction, then using above-mentioned condition formation urea, obtains IV-4.

Plan V I

Reagent:(a) acid chloride, K₂CO₃, dioxane/H₂O, and dicyclohexyl (2', 4', 6'- triisopropyl-[1,1'- joins Benzene] -2- bases) phosphine

Using Buchwald-Hartwig amination conditions, urea IV-3 chlorine function is optionally replaced by alkylamine Group, obtains (VI-1).Buchwald-Hartwig reactions are usually using palladium (0) catalyst such as Pd (PPh₃)₄With large volume Bronstead alkali such as KOt-Bu or KHMDS, are reacted in the mixture comprising ether solvents such as DME, dioxanes or THF. The method of palladium mediated amine coupling is described in Hartwig, J.F. (1998), Transition Metal Catalyzed Synthesis of Arylamines and Aryl Ethers from Aryl Halides and Triflates:Scope and Mechanism,Angew.Chem.Int.Ed.37:2046–2067。

Compound characteristic and property

In an exemplary embodiment, therapeutic compound may pass through the cytoplasmic membrane of cell.For example, compound There can be at least about 20%, 50%, 75%, 80%, 90% or 95% cell-permeability.

Compound as described herein can also have one or more following properties：The compound can substantially to cell or Subject is non-toxic；The compound 1000amu or can be less than for organic molecule or with 2000amu or less than 2000amu 1000amu small molecule；The compound can under ordinary atmospheric conditions have at least about 30 days, 60 days, 120 days, 6 months or The half-life period of 1 year；The compound can have the half-life period of at least about 30 days, 60 days, 120 days, 6 months or 1 year in solution； The compound can in solution, stable compared with resveratrol at least about 50%, 2 times, 5 times, 10 times, 30 times, 50 times or 100 times； The compound can promote DNA reparative factors Ku70 deacetylated effect；The compound can promote RelA/p65's de- Acetylating effect；The compound can increase general conversion ratio, and enhancing cell to the sensitivity of the TNF- Apoptosis induced Property.

In some embodiments, the compound of regulation Sirtuin is in (such as internal) effectively regulation silence regulation Under the concentration of the deacetylase activity of albumen, without suppression I classes histone deacetylase (HDAC), and/or ii Class HDAC any physical capacity.For example, in preferred embodiments, the compound of regulation Sirtuin is adjusted to be a kind of The compound of Sirtuin is saved, its is chosen had to activating the deacetylase activity of Sirtuin EC₅₀It is worth the EC than suppressing HDAC I and/or HDAC II₅₀Value it is small at least 5 times, and even preferably it is small at least 10 times, small 100 times or It is small or even 1000 times.Method for analyzing HDAC I and/or HDAC II activity is known in the art, and carries out such survey Fixed kit can in the market buy.See, for example, BioVision, Inc. (Mountain View, CA；Network address ) and Thomas Scientific (Swedesboro, NJ biovision.com；Network address thomassci.com).

In some embodiments, the compound of regulation Sirtuin is without regulation Sirtuin homologue Any physical capacity.In some embodiments, the activator of people's Sirtuin, in (such as in vivo) effective activation Under the concentration of the deacetylase activity of people's Sirtuin, lower eukaryotes may be derived from without activation, particularly Any physical capacity of the Sirtuin of yeast or human pathogen.For example, the compound of regulation Sirtuin can be through Select the EC for activating people's Sirtuin (such as SIRT1 and/or SIRT3) deacetylase activity having₅₀Value Than the EC of activated yeast Sirtuin (such as Sir2 (as Mycotoruloides, saccharomyces cerevisiae))₅₀Value is small at least 5 times, and very To it is more preferably small at least 10 times, small 100 times or even small 1000 times.In another embodiment, it is (special derived from lower eukaryotes Be not yeast or human pathogen) Sirtuin inhibitor, (for example in vivo) effectively suppress derive from low eucaryon Under the concentration of the deacetylase activity of biological Sirtuin, without times for suppressing the Sirtuin derived from people What physical capacity.For example, the inhibitory compound of Sirtuin can be chosen have for suppress people's silence regulation The IC of the deacetylase activity of albumen (such as SIRT1 and/or SIRT3)₅₀Value is than suppressing yeast Sirtuin (for example Sir2 (such as Mycotoruloides, saccharomyces cerevisiae)) IC₅₀Value it is small at least 5 times and even more preferably small at least 10 times, small 100 times or It is even small 1000 times.

In some embodiments, the compound of regulation Sirtuin can have the one or more silence regulations of regulation Protein homologue, such as one or more people SIRT1, SIRT2, SIRT3, SIRT4, SIRT5, SIRT6 or SIRT7 ability. In some embodiments, the compound of regulation Sirtuin has the ability of regulation SIRT1 and SIRT3 albumen.

In other embodiments, SIRT1 conditioning agents are in (such as in vivo) effectively regulation people SIRT1 deacetylase Under the concentration of activity, without other Sirtuin homologues are adjusted, such as one or more people SIRT2, SIRT3, SIRT4, SIRT5, SIRT6 or SIRT7 any physical capacity.For example, the compound of regulation Sirtuin can be chosen The ED for adjusting people's SIRT1 deacetylase activities being had₅₀Value people SIRT2s more one or more than regulation, SIRT3, SIRT3, SIRT4, SIRT5, SIRT6 or SIRT7 ED₅₀Value is small at least 5 times and even more preferably small at least 10 times, small 100 times Or even small 1000 times those compounds.In some embodiments, SIRT1 conditioning agents are without regulation SIRT3 albumen Any physical capacity.

In other embodiments, SIRT3 conditioning agents are in (for example, in vivo) effectively regulation people SIRT3 deacetylase The concentration of activity without the other Sirtuin homologues of any regulation (such as people SIRT1, SIRT2, SIRT4, SIRT5, One or more of SIRT6 or SIRT7) physical capacity.For example, the compound of regulation Sirtuin can be chosen institute The ED for adjusting people's SIRT3 deacetylase activities having₅₀Value people SIRT1s more one or more than regulation, SIRT2, SIRT4, SIRT5, SIRT6 or SIRT7 ED₅₀Value is small at least 5 times and even more preferably small at least 10 times, small 100 times or even It is small 1000 times.In one embodiment, any physical capacity of the SIRT3 conditioning agents without regulation SIRT1 albumen.

In some embodiments, binding affinity of the compound of regulation Sirtuin for Sirtuin It can be about 10^-9M、10^-10M、10^-11M、10^-12M or following.The compound for adjusting Sirtuin can be by Sirtuin pair In its substrate or NAD⁺The apparent K of (or other co-factors)_mValue reduce (activator) or increase (inhibitor) up at least about 2,3,4, 5th, 10,20,30,50 or 100 times.In some embodiments, K_mValue is determined using mass spectral analysis as described herein.It is preferred that work Change compound and reduce Sirtuin for its substrate or the K of co-factor_mThe degree of value compared with by resveratrol in similar concentration Lower caused degree is big, or reduces Sirtuin for its substrate or the K of co-factor_mValue reach by resveratrol compared with Caused K under low concentration_mValue.The compound of regulation Sirtuin can make the V of Sirtuin_maxValue increase is at least About 2,3,4,5,10,20,30,50 or 100 times.Adjust the compound regulation SIRT1 and/or SIRT3 albumen of Sirtuin Deacetylase activity ED₅₀Be worth less than about 1nM, less than about less than about 10nM, 100nM, less than about 1 μM, less than about 10 μM, Less than about 100 μM, or from about 1-10nM, from about 10-100nM, from about 0.1-1 μM, from about 1-10 μM or from about 10-100 μM.Adjust Save Sirtuin compound can adjust the deacetylase activity of SIRT1 and/or SIRT3 albumen up at least about 5,10, 20th, 30,50 or 100 times, it is that analysis by cell analysis or based on cell is determined.It is white relative to same concentrations Veratryl alcohol, the compound of regulation Sirtuin can induce the deacetylase activity of Sirtuin up at least about 10%th, 30%, 50%, 80%, 2 times, 5 times, 10 times, 50 times or 100 times.Adjust the compound regulation of Sirtuin SIRT5 ED₅₀It is worth the ED than adjusting SIRT1 and/or SIRT3₅₀Value is big at least about 10 times, 20 times, 30 times, 50 times.

Exemplary purposes

In some respects, the present invention is provided to adjust the level of Sirtuin and/or the method and purposes of activity, And its application method.

In some embodiments, the present invention provides the method and purposes of the compound using regulation Sirtuin, Wherein adjust Sirtuin compound activating Sirtuin, for example increase Sirtuin level and/or Activity.The compound for increasing the level of Sirtuin and/or the regulation Sirtuin of activity can be used for various treatments Using, including for example increase cell survival, and treatment and/or prevent many kinds of diseases and illness, including for example with aging or should Swash relevant disease or obstacle, diabetes, obesity, neurodegenerative disease, angiocardiopathy, blood clotting illness, inflammation, cancer And/or flush etc..The method and use of the present invention are included the compound of the regulation Sirtuin of pharmaceutical effective amount, example The compound for such as adjusting Sirtuin gives patient in need thereof.

Theory is not intended to be limited to, but believes the same area (example that activator of the invention can be in Sirtuin Such as, active position or the K of the active position is influenceed_mOr V_maxPosition) with Sirtuin interact.Think this i.e. one The reason for why activator and inhibitor of the Sirtuin of a little types can have substantial structure similitude.

In some embodiments, the compound of regulation Sirtuin specifically described herein can be used alone or and its Its compound is applied in combination.In some embodiments, can be by the compound of two or more regulation Sirtuins Mixture gives subject in need thereof.In another embodiment, the level of increase Sirtuin and/or work The compound of the regulation Sirtuin of property can be administered together with one or more following compounds：Resveratrol, palas flower Element, fisetin, piceatannol (piceatannol) or Quercetin.In an exemplary embodiment, increase silence is adjusted Save the compound and nicotinic acid or niacinamide riboside combination medicine-feeding of the level of albumen and/or the regulation Sirtuin of activity. In another embodiment, the compound of the level of Sirtuin and/or the regulation Sirtuin of activity is reduced It can be administered together with one or more following compounds：Niacinamide (NAM), shura are peaceful (suranim)；A kind of NF023 (G- albumen Antagonist)；NF279 (a kind of purinergic receptor antagonists)；Tuo Luosuo (Trolox) (6- hydroxyl -2,5,7,8- tetramethyls chroman - 2- formic acid)；(-)-epigallocatechin (hydroxyl is located at position 3,5,7,3', 4', 5')；(-)-epigallocatechin does not have food Sub- acid esters (hydroxy position 5,7,3', 4', 5', and gallate is located at position 3)；Cyanidin chloride (3,5,7,3', 4'- five Hydroxy chloride Huang (flavylium) salt)；Delphinidin chloride (3,5,7,3', 4', 5'- hexahydroxy chlorination Huang salt)；Red bayberry Flavones (cannabiscetin；3,5,7,3', 4', 5'- quercetagetin)；3,7,3', 4', 5'- pentahydroxyflavone；Gossypetin (3,5, 7,8,3', 4'- quercetagetin), Se Tingnuo (sirtinol)；And Si Tuo meter Xin (splitomicin)..In another embodiment party In case, the compound of one or more regulation Sirtuin can be used to treat or prevent various diseases with one or more (including for example cancer, diabetes, neurodegenerative disease, angiocardiopathy, blood clotting, inflammation, flush, obesity, aging, should Swash) therapeutic agent be administered together.In multiple embodiments, the combination of the compound comprising regulation Sirtuin Therapy can relate to the compound of (1) comprising one or more regulation Sirtuins with one or more therapeutic agents (for example, one Kind or a variety of described therapeutic agents in this article) pharmaceutical composition；And (2) one or more regulation Sirtuin Compound and the co-administered of one or more therapeutic agents, wherein the compound and therapeutic agent of the regulation Sirtuin are not Prepare in same composition (but may be present in identical kit or packaging, such as blister package or other multi-compartment bags In dress；Be present in can voluntarily be separated by user it is being connected, respectively in sealed container (such as tinfoil paper pouch)；Or exist In in the kit that wherein described compound is in the container separated with other therapeutic agents).When using separated preparation, adjust Saving the compound of Sirtuin can simultaneously, intermittently, staggeredly, before it, after which enter with the administration of another therapeutic agent OK, or in such ways combination is carried out.

In some embodiments, using compound as described herein to mitigate, prevent or treat disease or the side of obstacle Method and purposes may also comprise the egg of increase Sirtuin (such as people SIRT1, SIRT2 and/or SIRT3, or its homologue) White level.Increase protein level can be by being introduced into cell and complete by the one or more copy nucleic acids for encoding Sirtuin Into.For example, can be increased by the way that the nucleic acid for encoding Sirtuin is introduced into mammalian cell in mammalian cell The level of Sirtuin, such as by the way that coding GenBank to be logged in the amino acid sequence shown in numbering NP_036370 Nucleic acid introduces to increase SIRT1 level, and/or by the way that coding GenBank to be logged in the amino shown in numbering AAH01042 The nucleic acid of acid sequence introduces to increase SIRT3 level.

It is introduced into cell to increase the nucleic acid codified and Sirtuin of Sirtuin level (for example SIRT1 and/or SIRT3 albumen) sequence at least about 80%, 85%, 90%, 95%, 98% or 99% identical albumen.Example Such as, the nucleic acid for encoding the albumen can be with coding SIRT1 (such as GenBank log in numbering NM_012238) and/or SIRT3 is (for example GenBank log in numbering BC001042) albumen nucleic acid at least about 80%, 85%, 90%, 95%, 98% or 99% it is identical.Core Acid is alternatively and is preferable under stringent hybridization condition and encoding wild type Sirtuin (such as SIRT1 and/or SIRT3 albumen) Nucleic acid hybridization nucleic acid.Stringent hybridization condition may include in 0.2 × SSC in hybridizing and clean at 65 DEG C.When using Coding albumen (for example its for wild type Sirtuin fragment albumen) it is different from wild type Sirtuin During nucleic acid, the albumen is preferably biological activity, for example, can carry out deacetylated effect.Only need to express in cell A part for Sirtuin with biological activity.For example, the open country with logging in numbering NP_036370 with GenBank Albumen different raw type SIRT1, preferably comprises its core texture.The core texture is sometimes referred to as GenBank and logs in numbering NP_ 036370 amino acid 62-293, it is encoded by the GenBank nucleotides 237 to 932 for logging in numbering NM_012238, and it is included NAD is combined and substrate-binding domain.SIRT1 Core domain can also refer to：GenBank logs in numbering NP_036370's About amino acid 261 to 447, coded by its nucleotides 834 to 1394 for logging in numbering NM_012238 as GenBank； GenBank logs in numbering NP_036370 about amino acid 242 to 493, and it is logged in numbering NM_012238 core by GenBank Coded by thuja acid 777 to 1532；Or GenBank logs in numbering NP_036370 about amino acid 254 to 495, its by GenBank is logged in coded by numbering NM_012238 nucleotides 813 to 1538.Egg can be determined according to methods known in the art Whether retain biological function in vain, such as deacetylated ability.

In some embodiments, using regulation Sirtuin compound with mitigate, prevent or treat disease or The method and purposes of obstacle may also comprise reduction Sirtuin (such as people SIRT1, SIRT2 and/or SIRT3, or its homology Thing) protein level.Reduction Sirtuin level can be realized according to methods known in the art.For example, can be in cell Express siRNA, antisensenucleic acids or the ribozyme of targeted silent regulatory protein.Dominant negative (dominant can also be used Negative the mutant of Sirtuin), for example, can not carry out deacetylated mutant.For example, warp can be used It is described in such as Luo et al. (2001) Cell 107：SIRT1 mutant H363Y in 137.Or, it can be used and suppress transcription work Reagent.

Method and purposes for adjusting Sirtuin level also include the gene of regulation coding Sirtuin Transcription method and purposes, the corresponding mRNA of stabilisation/stabilization removal method and purposes, and it is known in the art its Its method and purposes.

Aging/stress

In one aspect, the present invention provides through by cell with the present invention increase Sirtuin level and/or The compound contact of the regulation Sirtuin of activity, so as to extend cell survival, increase ability of cell proliferation, slow down cell Aging, promote cell survival, delay cell ageing, simulation heat restriction effect, increase cell to stress resistance or prevent thin The method of born of the same parents' apoptosis.In an exemplary embodiment, the method and use of the present invention are included cell and regulation silence The compound contact of regulatory protein.

Method described herein and purposes can be used for increase cell, particularly blastema (to produce from organism, such as people Cell) can be kept in cell culture survival time.Embryonic stem cell (ES) and multipotential cell (pluripotent Cell the level and/or the regulation Sirtuin of activity for increasing Sirtuin can also) and by the cell of its differentiation be used Compound handle so that cell or its filial generation keep longer time in culture.Such cell can also be used for for example It is implanted into after modification (ex vivo modification) in vitro is carried out in subject.

In one aspect, the change of the level of increase Sirtuin and/or the regulation Sirtuin of activity can be used The cell that compound processing will be preserved for a long time.Cell may be present in suspension (such as haemocyte, serum, biological growth culture Base etc.) in, or be present in tissue or organ.For example, can be sunk with the regulation of the level and/or activity of increase Sirtuin The blood of the feeding blood from individual is collected in the compound processing of silent regulatory protein, haemocyte is preserved longer time.In addition, The change of the level of increase Sirtuin and/or the regulation Sirtuin of activity can also be used for legal medical expert's purpose blood Compound is preserved.Other cells that can extend its life-span through handling or protect it to anti-apoptotic include the cell example for consumption Such as derive from the cell (such as meat) of non-human mammal, or plant cell (such as vegetables).

Also increase Sirtuin can be applied in the development of mammal, plant, insect or microorganism and growth period The compound of the regulation Sirtuin of level and/or activity, for example to change, slow down or accelerated development and/or grow Journey.

In another aspect, the chemical combination of the regulation Sirtuin of the level of increase Sirtuin and/or activity Thing can be used for processing outstanding for transplanting or the cell of cell therapy, including such as solid tissue graft, organ graft, cell Supernatant liquid, stem cell, bone marrow cell etc..Cell or tissue can be autograft, allograft (allograft), same Plant isograft (syngraft) or xenograft.Can be by cell or tissue before administration/implantation, in administration/plant While entering, and/or after administration/implantation subject, handled with the compound of regulation Sirtuin.Can will be thin Born of the same parents or be organized in take out cell from provider's individual before, the cell or tissue (ex in vitro after being taken out from provider's individual Vivo) or after implantation acceptor handled.For example, the compound of available adjustment Sirtuin to provider or Acceptor individual carries out systemic processing, or level and/or the regulation silence regulation egg of activity with increase Sirtuin White compound on intracellular/tissue subgroup (subset) carries out locality processing.In some embodiments, can be additionally with another A kind of therapeutic agent for being used to extend graft survival, handles cell such as immunodepressant, cell factor, angiogenesis factor Or tissue (or provider/recipient's individual).

In other embodiments, it can be sunk in vivo with the regulation of the level and/or activity of increase Sirtuin The compound processing cell of silent regulatory protein, for example to increase its life-span or prevent Apoptosis.For example, can be heavy by using increase The compound of the level of silent regulatory protein and/or the regulation Sirtuin of activity is protected to handle skin or epidermal cell Skin is with anti-aging (for example, forming wrinkle, forfeiture elasticity etc.).In an exemplary embodiment, by skin with comprising Increase the pharmaceutical composition or cosmetics of the compound of the level of Sirtuin and/or the regulation Sirtuin of activity Composition is contacted.The exemplary skin disease or skin disorder that can be handled according to methods described herein and purposes, including with inflammation, Sunburn or naturally-aged correlation or the illness or disease that are induced by it.For example, composition can be used for preventing or treating contact skin It is scorching (including irritant contact dermatitis and allergic contact dermatitis), atopic dermatitis (also referred to as allergic dermatitis allergic eczema), photochemical Property keratosis, keratinization illness (including eczema), epidermolysis bullosa sick (including pemphigus), exfoliative dermatitis, seborrheica Dermatitis, erythema (including erythema multiforme and erythema nodosum), the damage as caused by Exposure to Sunlight or other light sources, lupus erythematosus discoides, Dermatomyositis, psoriasis, cutaneum carcinoma and naturally-aged effect.In another embodiment, the level of Sirtuin is increased And/or the compound of the regulation Sirtuin of activity can be used for wound to handle and/or burn to promote healing, including for example Once, two degree or third-degree burn and/or thermal burn, chemical burn or electric burn.Preparation can be partly administered to skin or viscous Membrane tissue.

The chemical combination of the regulation Sirtuin of level and/or activity comprising one or more increase Sirtuins The topical formulations of thing, it is also possible to be used as preventative (such as chemopreventive) composition.When being used in chemopreventive method When, occur the skin of the preceding processing easy infection of visible illness in specific individual.

The compound for adjusting Sirtuin can topically or systemically be delivered to subject.In an embodiment In, by injection, topical formulations etc., the compound for adjusting Sirtuin is partly administered to the tissue or device of subject Official.

In another embodiment, the regulation Sirtuin of the level of increase Sirtuin and/or activity Compound can be used for treating or preventing because of cell ageing induces or aggravated in subject disease or obstacle；For lower by The method and purposes of examination person's aging rate (such as after senescing)；Method and purposes for extending subject's life-span； The disease relevant with the life-span or the method and purposes of obstacle for treating or preventing；For treatment or prevention and ability of cell proliferation Relevant disease or the method for obstacle and purposes；And for treating or preventing disease or barrier caused by cellular damage or death The method and purposes hindered.In some embodiments, this method is not by the diseases for lowering those shortening subject's life-spans Incidence and work.In some embodiments, this method not by lower as the cause caused by disease (such as cancer) Dead rate and work.

In another embodiment, in order to increase the cell survival of subject in general manner and in order to protect its cell To resisting stress and/or to the purpose of anti-apoptotic, the regulation silence of the level and/or activity that increase Sirtuin can be adjusted The compound of albumen gives subject.It can trust that with compounds for treating subject as described herein, being passed through similar to subject is made Go through hormesis (hormesis) i.e., it is beneficial to organism and gently stress for its life-span can be extended.

The compound of the level for increasing Sirtuin and/or the regulation Sirtuin of activity can be given tested Person, with pre- anti-aging and relevant with aging consequence or disease, such as apoplexy, heart disease, heart failure, arthritis, hypertension And Alzheimer disease.Other illnesss being treated include eye diseases, such as ocular disorders relevant with eyes aging, for example in vain Cataract or glaucoma, glaucoma and macular degeneration.Also can be by the level and/or the regulation Sirtuin of activity that increase Sirtuin Compound give subject, for treating the disease relevant with cell death, such as chronic disease, to protect cell not dead Purpose.Exemplary diseases include those and hindered with nerve cell death, neuron dysfunction or muscle cell death or function Hinder relevant disease, such as Parkinson's, Alzheimer disease, multiple sclerosis, amyotrophic lateral sclerosis (amniotropic lateral sclerosis) and muscular dystrophy；AIDS；Fulminant hepatitis；The disease related to brain degeneration Disease, such as creutzfeldt-jakob disease (Creutzfeldt-Jakob disease), retinitis pigmentosa and cerebellar degeneration；Spinal cord development is not Good such as alpastic anemia；Ischemic disease such as miocardial infarction and apoplexy；Hepatopathy such as alcoholic hepatitis, hepatitis B With hepatitis C；Joint disease such as osteoarthritis；Atherosclerosis；Alopecia；The skin injury as caused by UV light；Flat tongue Moss；Atrophoderma；Cataract；And graft rejection.Cell death by operation, drug therapy, chemical contact or can also be put Penetrate contact caused.

Also the compound of the level for increasing Sirtuin and/or the regulation Sirtuin of activity can be given and suffers from There is an acute illness, for example the subject of organ or tissue's injury, such as the subject with apoplexy or miocardial infarction, or with ridge The subject of marrow damage.The compound for increasing the level of Sirtuin and/or the regulation Sirtuin of activity also may be used For repairing alcoholic liver (alcoholic ' s liver).

Angiocardiopathy

In another embodiment, a kind of method that the present invention provides treatment and/or prevention of cardiovascular disease, it passes through The compound of the level for increasing Sirtuin and/or the regulation Sirtuin of activity is given in need tested Person.

Can be used increase Sirtuin level and/or activity regulation Sirtuin compounds for treating or The angiocardiopathy of prevention, including cardiomyopathy or myocarditis, such as idiopathic cardiomyopathy, metabolic cardiomyopathy, alcoholic-toxic Cardiomyopathy, ischemic cardiomyopathy and hypertensive cardiomyopathy that cardiomyopathy, medicine trigger.Can also be used compound described herein with The disease that method and purposes are treated or prevented is dynamic for such as sustainer, coronary artery, arteria carotis, cerebrovascular arteries, the arteria renalis, ilium Arteries and veins, femoral artery are Ji the atherosclerotic condition (macrovascular diseases) of the Major Vessels such as popliteal artery (popliteal arteries).Its The vascular diseases that it can be treated or prevent include those and platelet aggregation, retinal arterioles, glomerulus parteriole (glomerular arteriole), vasa nervorum (vasa nervorum), heart parteriole, and eye, kidney, heart with Maincenter and the related vascular diseases of the related capillary bed of peripheral nervous system.Increase Sirtuin level and/or The compound of the regulation Sirtuin of activity can also be used for increasing the HDL levels in individual blood plasma.

The level and/or the compounds for treating of the regulation Sirtuin of activity for increasing Sirtuin can be used Other illnesss include ISR (such as after percutaneous coronary intervention), and with high density and low density cholesterol abnormal level Relevant illness.

In some embodiments, the regulation Sirtuin of the level of increase Sirtuin and/or activity Compound can be administered as a part for combination treatment together with another cardiovascalar agent.In some embodiments, increase Plus the level of Sirtuin and/or the compound of regulation Sirtuin of activity can be as one of combination treatment Divide and be administered together with antiarrhythmics agent.In another embodiment, the level and/or activity of Sirtuin are increased The compound of regulation Sirtuin can be administered as a part for combination treatment together with other cardiovascalar agents.

Cell death/cancer

Increasing the compound of the level of Sirtuin and/or the regulation Sirtuin of activity can give recently Receive or may will receive the subject of doses radiation or toxin.In one embodiment, radiation or toxin are received Dosage is for example administered as a part for work relative program or medical procedures as precautionary measures.In some embodiments, Not inadvertently receive radiation or the exposure of toxin.In such a situation it is preferred to the compound be administered as early as possible after exposure, to press down Apoptosis processed and follow-up developments are into acute radiation syndrome.

The compound of regulation Sirtuin can be also used for treatment and/or pre- anti-cancer.In some embodiments, The compound of the level of the increase Sirtuin and/or the regulation Sirtuin of activity can be used for treatment and/or pre- Anti-cancer.The reduction that energy limits the incidence of disease of age-related disease (such as cancer) establishes contact.Correspondingly, The increase of the level and/or activity of Sirtuin is advantageously used for treating and/or prevents age-related disease (such as cancer Disease) the incidence of disease.Can be brain and kidney using the exemplary cancer for the compounds for treating for adjusting Sirtuin Cancer；The cancer of hormone-dependence, including breast cancer, prostate cancer, carcinoma of testis and oophoroma；Lymthoma and leukaemia.With reality In the relevant cancer of body tumour, modulating compound can be directly given into the tumour.Cancer (such as white blood of blood cell Disease) can be by the way that modulating compound be administered in blood flow or treated in marrow.Also benign cell growth example can be treated Such as wart.The Other diseases that can be treated include autoimmune disease, such as systemic lupus erythematosus, chorionitis and arthritis, its In autoimmunity cell should be removed.Also the compounds for treating and virus infection (example of Sirtuin can be adjusted by being administered Such as bleb, HIV, adenovirus and HTLV-1) relevant pernicious and benign conditions.Or, cell can be obtained from subject, through external Processing gives identical or different subject to remove some undesirable cells (such as cancer cell), and return again to.

Also chemotherapeutant and the modulating compound described herein with active anticancer (can for example be induced Apoptosis Compound, subtract short-life compound or make cell to the compound of stress sensitive) co-administered.Chemotherapeutant in itself may be used Induce cell death with described herein or shorten the compound of life-span or increase to the regulation Sirtuin of stress sensitive It is applied in combination, and/or is used with other chemotherapeutic combinations.In addition to conventional chemotherapeutics, regulation as described herein The compound of Sirtuin also can be used together and not wished with suppressing to cause with antisense RNA, RNAi, or other polynucleotides Hope the expression of the cellular component of cell propagation.

The combination treatment of compound and conventional chemotherapeutics comprising regulation Sirtuin may be better than this area Known combination treatment, because the combination can make conventional chemotherapeutics play more large effect at lower doses.In preferred reality Apply in scheme, when the compound with adjusting Sirtuin is combined in use, for chemotherapeutant or conventional chemotherapy The combination of agent, effective dose (ED₅₀) than the ED of the single chemotherapeutant₅₀Small at least 2 times and even more preferably small 5 times, Small 10 times or even small 25 times.Conversely, when the compound with regulation Sirtuin described herein is combined in use, for this For the therapeutic index (TI) of the combination of class chemotherapeutant or such chemotherapeutant, it is than single conventional chemotherapy side The TI of case is high at least 2 times, and even more preferably still high 5 times, high 10 times or even high 25 times.

Neuronal disease/illness

In some respects, the compound of the regulation Sirtuin of the level of increase Sirtuin and/or activity Available for wound of the treatment with neurodegenerative disease, and central nervous system (CNS), spinal cord or peripheral nervous system (PNS) Or the patient of mechanical injuries.Neurodegenerative disease is usually directed to the reduction of human brain quality and volume, and it may be thin due to brain Born of the same parents' atrophy and/or it is dead caused by, the reduction of this human brain quality and volume than Healthy People caused by aging is more notable.Nerve Degenerative disease can be after long-term normal brain activity function be performed, due to progressive degeneration (such as nerve cell function in specific brain regions region It is not enough and dead) and be gradually in progress.On the other hand, neurodegenerative disease can rapid onset, such as those and wound or toxin phase Relevant neurodegenerative disease.The actual breaking-out that brain is degenerated may be compared with clinical expression early many years.The example of neurodegenerative disease Including (but being not limited to) Alzheimer disease (AD), Parkinson's (PD), Huntington disease (HD), amyotrophic lateral sclerosis (ALS；Luo Jieli Graves diseases (Lou Gehrig ' s disease)), dispersivity lewy body disease (diffuse Lewy body Disease), Chorea-acanthocytosis, primary lateral sclerosis, eye diseases (ophthalmoneuritis), phase chemotherapy induced DPN (for example originating from vincristine, taxol, bortezomib), the DPN of diabetes-induced and Buddhist Reed rely uncommon common Ji imbalance.The compound for increasing the level of Sirtuin and/or the regulation Sirtuin of activity can be used for treating this A little illnesss and as described below other illnesss.

AD is to cause the loss of memory, abnormal behaviour, personality change and the CNS illnesss of elaborative faculty decline.These lose with Link between certain types of brain cell death and brain cell is relevant with its supporting network (such as Deiter's cells) damage. Earliest period symptom includes losing nearest memory, false judgment and personality change.Body kinematics is uncontrolled, deadlock to cause by PD Firmly, tremble and dyskinesia CNS illnesss, the brain cell death prepared with brain in the region of dopamine associates.ALS (motions Neuronal disease) it is the CNS illnesss for attacking motor neuron (component for linking brain and skeletal muscle in CNS).

HD causes the uncontrolled, mental loss of motion and the neurodegenerative disease of emotional maladjustment to be another.Tay-Sachs disease (Tay-Sachs disease) and sandhoff disease (Sandhoff disease), are wherein GM2 gangliosides and beta-amino The associated sugars lipid substrate of hexoside enzyme (hexosaminidase) accumulates in nervous system and triggers acute neurodegenerative Glycolipid storage diseases (glycolipid storage diseases).

Well known, Apoptosis works in terms of the AIDS lesions of immune system.It can be used however, HIV-1 also induces The neurological disorder of the compounds for treating of the regulation Sirtuin of the present invention.

Neuron loss (neuronal loss) also for prion disease (prion diseases) (such as people's creutzfeldt-jakob disease, Ox BSE (rabid ox disease), sheep and the itch of goat disease and cat cat class spongiform encephalopathy (FSE)) prominent features.Increase silence The compound of the level of regulatory protein and/or the regulation Sirtuin of activity can be used for treating or preventing and be drawn by these diseases The neuron loss risen.

In another embodiment, the regulation Sirtuin of the level of increase Sirtuin and/or activity Compound can be used for treating or preventing any disease or obstacle for being related to aixs cylinder sick (axonopathy).Distal axonopathy is one Class is as the peripheral nerve disease produced by certain metabolism or toxicity entanglement of peripheral nervous system (PNS) neuron.It is right for nerve Metabolism or the disorderly most commonly response of toxicity, and therefore may be by metabolic disease such as diabetes, kidney failure, shortage syndrome Such as malnutritive and alcoholism, or as caused by the effect of toxin or medicine.Those patients with distal axonopathy are led to Symmetry gloves-socks sample sensation-dyskinesias is often presented.Also occur deep layer tendon reflex and autonomic nerves system in involved area (ANS) function is lost or reduced.

Diabetic neuropathy is the nervous disorders relevant with diabetes.Relevant with diabetic neuropathy is more normal See that illness is benumbed including third nerve；Mononeuropathy；Mononeuritis multiplex；Diabetes impotency；Painful is more Neuropathy；Autonomic neuropathy；And ventral thoracic nerve disease.

Peripheral nerve disease is changed into the medical terminology for the neurotrosis to peripheral nervous system, and it is probably by neural disease Disease or as caused by the side effect of systemic disease.The main cause of peripheral neuropathy include epileptic attack, nutritional deficiency and HIV, but diabetes are most probable reasons.

In an exemplary embodiment, the regulation silence of the level and/or activity that increase Sirtuin is adjusted The compound for saving albumen can be used for treating or preventing multiple sclerosis (MS), including recurrent MS and monosymptom MS, and other de- Myelin illness (demyelinating condition), such as chronic inflammatory Demyelinating Polyneuropathy disease are (CIDP) or associated Illness.

In still another embodiment, the regulation Sirtuin of the level of increase Sirtuin and/or activity Compound can also be used for treating traumatic nerve injury, including because disease, damage (including operation intervention) or environment wound are (such as refreshing Through toxin, alcoholism etc.) caused by wound.

The compound of the level of increase Sirtuin and/or the regulation Sirtuin of activity can also be used for pre- Prevent, treat and alleviate the symptom of various PNS illnesss.Term " peripheral neuropathy " includes being located at outside brain and spinal cord of wide scope The illness that has been damaged of nerve (i.e. peripheral nerve).Peripheral neuropathy can also refer to peripheral neuritis, or if be related to perhaps During polyneural, term polyneuropathy or multiple neuritis can be used.

The level and/or the compounds for treating of the regulation Sirtuin of activity for increasing Sirtuin can be used PNS diseases include：Diabetes, leprosy, charcot-Marie-Tooth disease (Charcot-Marie-Tooth disease), Ge-bar Er Shi Syndrome (Guillain-Barr é syndrome) and wall neuropile neuropathy (Brachial Plexus Neuropathy) (neck and the first chest root, nerve cord, the disease of the peripheral nerve component of rope (cord) and brachial plexus).

In another embodiment, the compound of regulation Sirtuin can be used for treating or preventing polyglutamyl amine Disease.It is red that exemplary many glutamine diseases include spinobulbar muscular atrophy (Kennedy disease), Huntington disease (HD), dentate nucleus Core globus pallidus nucleus subthalamicus atrophy (Dentatorubral-pallidoluysian atrophy) (Hao crow river syndrome (Haw River syndrome)), 1 type spinocebellar ataxia, 2 type spinocebellar ataxias, 3 type Spinocerebellars be total to Ji imbalance (Ma-about sick (Machado-Joseph disease)), 6 type spinocebellar ataxias, 7 type Spinocerebellars are total to Ji imbalance and 17 type spinocebellar ataxias.

In some embodiments, the present invention provides treatment central nervous system cell to prevent because flowing in response to blood The method that cell lowers caused injury.The injury severity that can be generally prevented will largely depend on blood and flow to cell Decreased extent and reduction duration.In one embodiment, apoptotic or necrotic cell death can be prevented.Another In individual embodiment, the injury that ischemic can be prevented to be mediated, such as cytotoxic edema or central nervous system tissue's anoxemia Disease.In each embodiment, central nervous system cell can be cord cell or brain cell.

On the other hand include giving subject by the compound for adjusting Sirtuin, lacked with treating central nervous system Courageous and upright illness.There are many central nervous system ischemic conditions to be controlled by the compound of regulation Sirtuin as described herein Treat.In one embodiment, ischemic conditions are to cause any types ischemic central lesion, such as apoptotic Or the apoplexy of necrosis, cytotoxic edema or central nervous system tissue's anoxic.Apoplexy may influence brain to appoint What region, or as caused by any commonly known cause of disease that apoplexy can be caused to occur.In another selection of the embodiment, in Wind is brain stem apoplexy.In another selection of the present embodiment, apoplexy is small headstroke.In another selection of the present embodiment In, apoplexy is embolic stroke (embolic stroke).In another selection of the present embodiment, apoplexy is in hemorrhagic Wind.In other embodiments, apoplexy is embolic stroke.

On the other hand, the compound of regulation Sirtuin can be administered, with central nervous system ischemic conditions The infraction size of ischemic core is reduced afterwards.Moreover, the compound of regulation Sirtuin also can be valuably administered, with maincenter After nervous system ischemic conditions, the size of ischemic penumbra or transitional region is reduced.

In one embodiment, composition of medicine therapy may include be used for treat or prevent neurodegenerative or with The medicine or compound of the relevant secondary conditions of these illnesss.Therefore, composition of medicine therapy may include that one or more silences are adjusted Save the activator and one or more anti-neurodegeneration medicines of albumen.

Blood clotting illness

In other side, increase the compound of the level of Sirtuin and/or the regulation Sirtuin of activity Available for treatment or prevention blood clotting illness (or hemostasis illness).As being convertibly used for herein, term " hemostasis ", " blood Solidification " and " blood clotting " refer to that control is bled, including vessel retraction and the physiological property of condensation.Blood clotting assists to maintain to feed Circulation integrality of the newborn animal after injury, inflammation, disease, birth defect, dysfunction or other destructions (disruption). Moreover, the formation of clot not only limits bleeding (anastalsis) in the case of injury, it is also possible in atherosclerosis disease Aspect, causes serious organ injury and death because blocking important artery or vein.Therefore thrombus is in wrong time and place The blood clotting of formation.

Then, the present invention provides anti-agglomeration and curing thrombus, its object is to suppress blood clotting to be formed, to prevent or Treat blood clotting illness, such as miocardial infarction, apoplexy, limbs loss or pulmonary embolism caused by peripheral arterial disease.

The term " modulation hemostasis " convertibly used herein and " modulating hemostasis " include induction (for example stimulate or increase) Hemostasis, also including suppressing and (such as lowering or reduce) hemostasis.

On the one hand, the present invention provides through the level of administration increase Sirtuin and/or the regulation silence of activity The compound of regulatory protein, to lower or suppress the method stopped blooding in subject.The compositions disclosed herein, method and purposes Available for treatment or prevention thrombotic disease.Terms used herein " thrombosis illness " is included with excessive or undesirable Blood coagulation or styptic activity, or any illness or disease that hypercoagulable state is characterized.Thrombotic disease includes being related to Platelet adhesion reaction and thrombotic disease or obstacle, thereby increases and it is possible to be shown as formed as the possibility increase of thrombus, for example, form blood Bolt quantity increases, thrombosis, thrombotic family's sexual orientation occurs in low age period and thrombus shape occur in rare locationss Into.

In another embodiment, composition of medicine scheme may include to be used to treat or prevent blood clotting illness, or with The medicine or compound of the relevant secondary conditions of these illnesss.Therefore, composition of medicine scheme may include that one or more increases are heavy The compound of the level of silent regulatory protein and/or the regulation Sirtuin of activity and one or more anti-agglomerations or antithrombotic Form medicine.

Body weight control

On the other hand, the compound of the regulation Sirtuin of the level of increase Sirtuin and/or activity Available for the increased weight or obesity for treating or preventing subject.For example, increase Sirtuin level and/or activity The compound for adjusting Sirtuin can be used for for example treating or preventing genetic obesity, dietary obesity, hormone phase The obesity of pass, the obesity relevant with administration medicine, increase for mitigating subject's body weight or mitigation or prevention subject body weight Plus.Need such treatment subject can it is fat for it, be possible to become fat, overweight or be possible to become it is overweight by Examination person.Fat or overweight subject is likely to become, for example can be taken the photograph based on family history, science of heredity, diet, active level, medicine Take or its various combination is determined.

In other embodiments, the regulation silence of the level and/or activity that increase Sirtuin can be adjusted The compound of albumen give with it is various can be by the Other diseases and disease that promote the weight loss of subject and treat or prevent The subject of disease.Such disease includes such as hypertension, hyperpiesia, high blood cholesterol, dyslipidemia, type ii diabetes, pancreas Insulin resistance, poor glucose tolerance, hyperinsulinemia, coronary heart diseases and angina pectoris, congestive heart failure, apoplexy, courage knot Stone, cholecystitis and cholelithiasis, gout, osteoarthritis, obstructive sleep apnea and breathing problem, some types of cancer (examples Such as carcinoma of endometrium, breast cancer, prostate cancer and colon cancer), bad (for example menstruation is not or not complications of pregnancy, female reproductive health Regular, infertile, irregular ovulation), bladder control problem (such as stress urinary incontinence)；Uric acid renal lithiasis；Psychotropic disorders (example As melancholy, eating disorder disease, the bodily image of distortion and self-respect are reduced).Finally, the patient with AIDS can be directed to AIDS group Therapy response is closed, and develops into fat nutritional disorders or insulin resistance.

In another embodiment, the regulation Sirtuin of the level of increase Sirtuin and/or activity Compound can be used for suppressing Adipogenesis or Adipocyte Differentiation in vitro or in vivo.Such method and purposes can be used for treating Or prevention obesity.

In other embodiments, the regulation Sirtuin of the level of increase Sirtuin and/or activity Compound can be used for lowering appetite and/or increase satietion, therefore cause weight loss or avoid increased weight.Need such control The subject for the treatment of can be overweight, fat subject, or be possible to become overweight or fat subject.This method is included By doses (such as in pill (pill) form) once a day or every two days once or weekly give subject.Should Dosage can be " appetite attenuating dosage ".

In an exemplary embodiment, the regulation silence of the level and/or activity that increase Sirtuin is adjusted The compound of albumen is saved, can be administered with the combination treatment for treating or preventing increased weight or obesity.For example, can be by one Kind or it is a variety of increase Sirtuin level and/or activity regulation Sirtuin compound with one or more Antiadipositas drug combination medicine-feeding.

In another embodiment, the regulation silence of level and/or activity that increase Sirtuin can be administered is adjusted The compound of albumen is saved, to lower the increased weight triggered by medicine.For example, level and/or the work of increase Sirtuin The compound of the regulation Sirtuin of property, can with can stimulate appetite or cause increased weight (be particularly due to except Increased weight caused by factor beyond hydropexis) medicine be administered by combination treatment.

Metabolic disorder/diabetes

On the other hand, the compound of the regulation Sirtuin of the level of increase Sirtuin and/or activity Available for treatment or prevention metabolic disorder, such as insulin resistance, pre-diabetes, type ii diabetes, and/or its complication. Increasing the administration of the compound of the level of Sirtuin and/or the regulation Sirtuin of activity can increase subject's Insulin sensitivity and/or attenuating insulin level.The subject for needing such treatment can be with insulin resistance or II types Other premonitory symptoms of diabetes, with type ii diabetes or the subject for being possible to develop these any illnesss.For example, should be by Examination person can be with insulin resistance, such as with hyperinsulinism cyclical level and/or associated illness, such as hyperlipemia, Fatty dyspoiesis, hypercholesterolemia, poor glucose tolerance, elevated blood glucose levels, other performances of syndrome X, hypertension, The subject of atherosclerosis and lipodystrophia.

In an exemplary embodiment, the regulation silence of the level and/or activity that increase Sirtuin is adjusted The compound of section albumen can be administered with the combination treatment for treating or preventing dysbolism.For example, can be by one or more Increase the compound and one or more anti-diabetics of the level of Sirtuin and/or the regulation Sirtuin of activity Medicine combination medicine-feeding.

Inflammatory disease

On the other hand, the compound of the regulation Sirtuin of the level of increase Sirtuin and/or activity Available for the disease or obstacle treated or prevented with inflammation-related.Can in before inflammatory episode, breaking-out when or breaking-out after be administered increase Plus Sirtuin level and/or activity regulation Sirtuin compound.When preventive use, describedization Compound is administered preferably before any inflammatory reaction or symptom.The administration of the compound can prevent or weaken inflammatory reaction or disease Shape.

In another embodiment, the regulation Sirtuin of the level of increase Sirtuin and/or activity Compound can be used for treating or preventing allergy and respiratory condition, including asthma, bronchitis, pulmonary fibrosis, anaphylaxis Rhinitis, oxygen poisoning, pulmonary emphysema, chronic bronchitis, ARDS and any chronic obstructive pulmonary disease (COPD). The compound can be used for treatment virus infection, including hepatitis B and hepatitis C.

In addition, the compound of the level of increase Sirtuin and/or active regulation Sirtuin can be used for Treatment autoimmune disease, and/or the inflammation relevant with autoimmune disease, such as arthritis, including rheumatoid arthritis, Psoriasis arthropathica and ankylosing spondylitis, and organ-tissues autoimmune disease (such as Raynaud's syndrome (Raynaud's syndrome)), ulcerative colitis, Crohn disease, portacaval mucositis, chorionitis, myasthenia gravis, transplanting Repulsion, endotoxin shock, sepsis, psoriasis, eczema, dermatitis, multiple sclerosis, autoimmune thyroiditis, uvea Inflammation, systemic lupus erythematosus, Addision's disease (Addison's disease), the pluriglandular disease of autoimmunity (also referred to as itself are exempted from Epidemic disease polyglandular syndrome) and Graves disease.

In some specific embodiments, the regulation of the level and/or activity of one or more increase Sirtuin The compound of Sirtuin can be used alone, or be applied in combination with other compounds for treating or preventing inflammation.

Flush

On the other hand, the compound of the regulation Sirtuin of the level of increase Sirtuin and/or activity can Incidence or severity for lowering flush and/or hectic fever (hot flash) as the symptom of illness.For example, the side Method including the use of increase Sirtuin level and/or activity regulation Sirtuin compound, it is independent or with Other pharmaceutical agent combinations are used, to lower the flush of cancer patient and/or the incidence or severity of hectic fever.In other embodiments In, this method provide using increase Sirtuin level and/or activity regulation Sirtuin compound with Lower menopause and the flush and/or the incidence or severity of hectic fever of postmenopausal women.

On the other hand, the compound of the regulation Sirtuin of the level of increase Sirtuin and/or activity can Generation as the flush and/or hectic fever (for example, drug-induced flush) for lowering the side effect as another medicinal treatment The therapy of rate or severity.In some embodiments, the method for the flush induced for treatment and/or prophylactic agent includes will The regulation of compound containing at least one induction flush and the level and/or activity of at least one increase Sirtuin is sunk The preparation of the compound of silent regulatory protein gives patient in need.In other embodiments, induced for medicine The method of flush include, one or more compounds for inducing flush and one or more regulation silences regulation eggs are administered respectively White compound, for example wherein the compound of regulation Sirtuin is not formulated in same combination with inducing the medicine of flush In thing.When using separated preparation, the compound (1) for adjusting Sirtuin can simultaneously be given with inducing the medicine of flush Medicine, (2) are administered intermittently together with inducing the medicine of flush, and (3) stagger administration with inducing the medicine of flush, and (4) are in induction flush Drug administration before be administered, (5) are administered after the drug administration of flush is induced, and (6) are carried out with its various various combination Administration.Example sexflush induce medicine include such as nicotinic acid, Raloxifene (faloxifene), resist melancholy agent, antipsychotic drug, Chemotherapeutant, calcium channel blocker and antibiotic.

In one embodiment, the regulation Sirtuin of the level of increase Sirtuin and/or activity Compound can be used for the flush for reducing vasodilator or antilipemic (including hypercholesterolemia thing and lipotropic drug) Side effect.In an exemplary embodiment, increase the regulation silence regulation of the level and/or activity of Sirtuin The compound of albumen can be used for reducing the flush relevant with delivery of niacin.

In another embodiment, the present invention provides treatment and/or prevents hyperlipemia and lower flush side effect Method.In another representational embodiment, this method is including the use of the level and/or activity for increasing Sirtuin The compound of Sirtuin is adjusted, to reduce the flush side effect of Raloxifene.In another representational embodiment, This method is including the use of the level for increasing Sirtuin and/or the compound of the regulation Sirtuin of activity, to subtract The flush side effect of few antimelancholic or antipsychotic drug.For example, increase Sirtuin level and/or activity The compound of regulation Sirtuin can be used in conjunction with (separating with serotonin reuptake inhibithors or 5HT2 receptor antagonists Or be administered together).

In some embodiments, the regulation Sirtuin of the level of increase Sirtuin and/or activity Compound can be used as a part for the treatment of serotonin reuptake inhibithors (SRI) to reduce flush.Another representational In embodiment, the compound for increasing the level of Sirtuin and/or the regulation Sirtuin of activity can be used for subtracting The flush side effect of few chemotherapeutant such as endoxan and TAM.

In another embodiment, the regulation Sirtuin of the level of increase Sirtuin and/or activity Compound can be used for reduce calcium channel blocker such as Amlodipine flush side effect.

In another embodiment, the regulation Sirtuin of the level of increase Sirtuin and/or activity Compound can be used for reduce antibiotic flush side effect.For example, the level and/or the tune of activity of increase Sirtuin The compound of section Sirtuin can be applied in combination with lavo-ofloxacin.

Ocular disorders

An aspect of of the present present invention is method for suppressing, lowering or treat vision impairment, and this method is by by therapeutic agent The conditioning agent of the Sirtuin selected from compound disclosed herein of amount or its pharmaceutically acceptable salt, prodrug or metabolism Derivative gives patient.

In terms of some of the present invention, vision impairment is due to caused by the injury to optic nerve or central nervous system. In specific embodiments, optic nerve injury is as caused by high intraocular pressure (such as the high intraocular pressure as caused by glaucoma). In other specific embodiments, by neural swelling, (it is often with infecting or being immunized (such as autoimmunity) instead for optic nerve injury Should be relevant (such as in optic neuritis)) it is caused.

In terms of some of the present invention, vision impairment is as caused by the injury of retina.In specific embodiments, depending on The injury of nethike embrane is caused by the obstacle (for example, atherosclerosis, vasculitis) in the blood flow for flowing to eyes.Specific real Apply in scheme, the injury of retina is to destroy (disruption of macula) (for example, exudative or nonexudativeage by macula lutea Macular degeneration) it is caused.

The disease of exemplary retina includes the related macular degeneration of Exudative Age, the macula lutea of nonexudativeage age correlation Denaturation, retina electronics the prosthese macular degeneration related with the RPE transplanting ages, acute many focus tabular pigment epitheliums are sick, acute Retinal necrosis, best's disease, branch retinal artery occlusion, branch retinal vein occlusion, cancer association and correlation from Body immunity retinopathy, CRAO, thrombosis of central vein of retina, Central Serous choroid view Film lesion (central serous chorioretinopathy), eales disease (Eales Disease), premacular membranes The denaturation of (epimacular membrane), dot matrix, huge aneurysm, diabetic macular edema, Ai Erwen-cover this macular edema New blood vessel film, dispersivity list under (Irvine-Gass Macular Edema), macula hole (macular hole), retina The subacute neuroretinitis in side, non-psaudophakic cystoid macular oedema (nonpseudophakic cystoid macular Edema), pseudo-tissue histoplasmosis syndrome (presumed ocular histoplasmosis syndrome), exudative Detachment of retina, postoperative detachment of retina, proliferative retinal disengaging, rhegmatogenous detachment of retina, traction property retina take off From, retinitis pigmentosa, the CMV retinitiss, retinoblastoma, Prematurity, shot shape retinopathy (birdshot retinopathy), background diabetic retinopathy, proliferating diabetic retinopathy, hemoglobin The husky retinopathy (Purtscher Retinopathy) of characteristic of disease retinopathy, pul, Wa Er Salva's retinopathies (Valsalva Retinopathy), juvenile retinoschisis (juvenile retinoschisis), senile view Film splitting disease, Tai Ersong syndromes (Terson Syndrome) and white point syndrome (white dot syndromes).

Other examples disease includes eye bacterium infection (for example, conjunctivitis, keratitis, tuberculosis, syphilis, gonorrhoea), disease Poison infection (such as eye herpes simplex virus (ocular herpes simplex Virus), varicellazoster virus, big and small The cellular virus retinitis, human immunodeficiency virus (HIV)) and progressive Outer Retinal Necrosis secondary to HIV or its Its HIV- is associated and other immune deficiencies-relevance eye diseases.In addition, eye diseases include fungal infection (such as monilial arteries and veins Network film inflammation (Candida choroiditis), histoplasmosis), protozoal infections (such as toxoplasmosis) and other diseases Case such as ocular toxocariasis and sarcoidosis.

An aspect of of the present present invention be for suppress, lower or treat with chemotherapeutic agent (for example, neurotoxicity medicine, Raise the medicine such as steroids of intraocular pressure) method of subject's vision impairment for the treatment of, it is by by the sheet of therapeutic dose The conditioning agent of Sirtuin disclosed in text gives the subject that such treatment needs.

Another aspect of the invention is for suppressing, lowering or treating (including the eye or other in prone position being performed the operation Operation, such as operation on spinal cord when) subject's vision impairment method, it is by by the disclosed herein heavy of therapeutic dose The conditioning agent of silent regulatory protein gives the subject that such treatment needs.Ocular operation includes cataract, irotomy and crystalline substance Shape body is replaced.

Another aspect of the invention is the eye diseases that treatment (including the suppress and prophylactic treatment) age is related, including it is white interior The method of barrier, xerophthalmia, age related macular degeneration (AMD), retinal damage etc., it is by by this paper institutes of therapeutic dose The conditioning agent of disclosed Sirtuin gives the subject that such treatment needs.

Another aspect of the invention is prevention or treat by stress, chemistry injure or the caused ocular damage of irradiation side Method, it by the conditioning agent of the Sirtuin disclosed herein of therapeutic dose by giving the tested of such treatment needs Person.Irradiation to eyes or electromagnetically damage may include as CRT or exposed to those damages caused by sunlight or UV.

In some embodiments, composition of medicine scheme may include to be used to treat or prevent ocular disorders or with these illnesss have The medicine or compound of the secondary conditions of pass.Therefore, composition of medicine scheme may include the work of one or more Sirtuins Agent and one or more therapeutic agents for being used to treat ocular disorders.

In some embodiments, can by the conditioning agent of Sirtuin with for reducing intraocular pressure therapy be combined into Row administration.In another embodiment, can by the conditioning agent of Sirtuin with for treating and/or preventing glaucoma Therapy is combined and is administered.In another embodiment, can by the conditioning agent of Sirtuin with for treating and/or preventing The therapy of optic neuritis is combined and is administered.In some embodiments, the conditioning agent of Sirtuin can be controlled with being used for Treat and/or the therapy combination of prevention CMV retinopathies is administered.In another embodiment, can be by Sirtuin Conditioning agent be administered with for treating and/or prevent the therapy of multiple sclerosis to be combined.

The relevant disease of mitochondria and obstacle

In some embodiments, the present invention is provided to treat because of the disease or obstacle that mitochondria activity increases and benefits Method and purposes.The compound that the method and use of the present invention include the regulation Sirtuin by upper effective dose is treated is given Give the subject of needs.Mitochondria activity increase refers to increase while mitochondrial total amount (such as mitochondrial quality) is maintained Plus therefore mitochondrial activity, the mitochondrial quantity that increases simultaneously increase mitochondrial activity (for example, by stimulating mitochondrial life Thing is generated), or its combination.In some embodiments, because mitochondria activity increases and the disease or obstacle that benefit, including with line The relevant disease of mitochondria function obstacle or obstacle.

In some embodiments, for treating because of the disease or the method for obstacle and use that mitochondria activity increases and benefits Way may include to determine the subject with mitochondria dysfunction.Method and purposes for diagnosing mitochondria dysfunction can be wrapped Include molecular genetics, pathology and/or biochemical analysis.The disease relevant with mitochondria dysfunction or obstacle are included so Disease and illness, wherein mitochondrial respiratory chain activity deficiency causes the pathologic, physiologic of such disease or obstacle in mammal Development.Because the disease or obstacle that mitochondria activity increases and benefits are generally comprised for example, being damaged by the oxidation of free radical mediated The disease of tissue degeneratiaon, cell inadequately carry out the disease of apoptosis caused by wound, and cell can not carry out the disease of apoptosis.

In some embodiments, the present invention is provided to treat because of the disease or obstacle that mitochondria activity increases and benefits Method and purposes, this method includes one or more compounds for adjusting Sirtuins and another therapeutic agent (example It such as can be used for the medicament for the treatment of mitochondria dysfunction, or available for the disease or obstacle for reducing and being related to mitochondria dysfunction The medicament of relevant symptom) combine and give subject in need.

In exemplary embodiment, the present invention is provided to treat because mitochondria activity increases and the disease that benefits or The method and purposes of obstacle, subject is given by the compound of the regulation Sirtuin by upper effective dose is treated.Example Property disease or obstacle include for example neuromuscular disorder (such as Buddhist Reed rely uncommon incoordination (Friedreich ' s Ataxia), Muscular dystrophy, multiple sclerosis etc.), neuron unstability illness (such as epileptic attack, antimigraine), hypoevolutism, In neurodegenerative disorders (such as Alzheimer disease, Parkinson's, amyotrophic lateral sclerosis), ischemic, renal tubule acid Poison, age related neurodegeneration and cognitive decline, chemotherapy are tired, the age is related or phase chemotherapy induced menopause or the moon Through cycle or ovulation are irregular, (such as calcium accumulation, excitotoxicity, nitric oxide are sudden and violent for mitochondrial myopathy, mitochondrial biogenesis Dew, anoxic etc.) and mitochondria imbalance.

Muscular dystrophy refers to a class and is related to the disease that myoneural histological structure deteriorates with function, and it often leads to skeletal muscle Atrophy and myocardial dysfunction, such as Duchenne muscular dystrophy (Duchenne muscular dystrophy).In some realities Apply in scheme, the compound of regulation Sirtuin can be used for the decay rates for lowering muscle function ability, and for strengthening Functional status with muscular dystrophy patient muscle.

In some embodiments, the compound of regulation Sirtuin can be used for treatment mitochondrial myopathy.Line grain Body myopathy scope is weak from the slight slow progressive of outer eye muscle, to serious fatal infancy myopathy and multisystem brain Myopathy (encephalomyopathy).Some syndromes are it has been determined that some are overlapping between them.Influence the establishment of muscle Syndrome include progressive ophthalmoplegia externa, Ka-plug syndrome (Kearns-Sayre syndrome) (have ophthalmoplegia, Pigmentary retinopathy, cardiac conduction defect, cerebellar ataxia and sensorineural hearing loss), MELAS syndrome (mitochondrias Property brain myopathy, lactic acidosis and apoplexy sample breaking-out (stroke-like episodes)), MERFF syndromes (myoclonia type Epilepsy, irregular red fiber (ragged red fibers)), limb girdle distribute weak (limb-girdle distribution ) and infantile myopathy (benign, severe with lethal) weakness.

In some embodiments, the compound of regulation Sirtuin can be used for treatment to be damaged by mitochondrial toxicity Evil, such as due to toxicity caused by calcium accumulation, excitotoxicity, nitric oxide exposure, drug-induced toxicity damage or anoxic The patient of infringement.

In some embodiments, the compound of regulation Sirtuin can be used for treating relevant with mitochondria imbalance Disease or obstacle.

Muscle performance

In other embodiments, the present invention is provided to strengthen the method and purposes of muscle performance, it is controlled by administration The compound of the regulation Sirtuin of effective dose in treatment.For example, the compound of regulation Sirtuin can be used for improving Body endurance (for example, carry out physical task such as moving, manual labor, motor activity), suppress or delay physical fatigue, Increase blood oxygen levels, the energy for the individual that improves health, strengthen ability to work and persistence, reduction muscular fatigue, reduce pressure, Strengthen heart and the lactic acid in cardiovascular function, improvement sexuality, increase muscle ATP levels and/or reduction blood.In some realities Apply in scheme, this method and purposes include a certain amount of increase mitochondria activity is administered, promote mitochondrial biological generation and/or Increase the compound of the regulation Sirtuin of mitochondrial mass.

Exercise performance refers to the ability that the muscle of sportsman is presented when participating in motor activity.The enhanced motion effect of institute Energy, intensity, speed and endurance, are by the increase of muscle contraction strength, the increase of contraction of muscle amplitude, are stimulating with shrinking Between the muscle response time shortening and measure.Sportsman refers to participates in motion to any degree, and seek can be at it The individual for promoting intensity, speed and endurance level, such as body builder (body builder), cycling are reached in function Member, long-distance runner, dash man etc..Enhanced sports performance efficiency by can overcome muscular fatigue ability, keep it is longer when Between vigor ability and shown with the ability more effectively taken exercise.

In the case of sportsman's muscle performance, expecting to produce allows to play or instruct under higher tolerant levels for a long time Experienced situation.

It is expected that the method and use of the present invention are also effective, including acute in terms of the related pathologic conditions for the treatment of muscle Sarcopenia (acute sarcopenia), for example muscular atrophy and/or with burn, bed, limbs ligamentopexis or chest, abdomen Portion and/or the relevant cachexia of plastic surgery major operation.

In some embodiments, the present invention provides the new dietary composition of the conditioning agent comprising Sirtuin, Its preparation method and using said composition to improve the method for sports performance efficiency.Then, the motion that the present invention is defined extensively to participation, People including needing work of the motion of endurance with needing repeated muscular movement, which provide to have, improves physical endurance and/or suppression Therapeutic composition, the bag and bottle of the effect of physical fatigue processed.Such dietary composition can additionally comprise electrolyte, coffee Cause, vitamin, carbohydrate etc..

Other purposes

Increase Sirtuin level and/or activity regulation Sirtuin compound can be used for treat or Pre- preventing virus infection (such as influenza virus, herpesviral or papilloma virus infection) is used as antifungal agent.In some realities Apply in scheme, increase the compound of the level of Sirtuin and/or the regulation Sirtuin of activity, group can be used as A part for composite medicine therapy is administered together with another therapeutic agent for treating viral disease.In another embodiment In, increase the compound of the level of Sirtuin and/or the regulation Sirtuin of activity, can be treated as composition of medicine A part for method is administered together with another antifungal agent.

The subject that as described herein can be treated includes eucaryote, such as mammal such as people, sheep class, ox Class, horse class, pig class, canine, cat class, non-human primates, mouse and rat.Accessible cell includes eukaryotic, for example From the cell of foregoing subject, or plant cell, yeast cells and prokaryotic such as bacterial cell.For example, can be by modulability Compound gives farm-animals, to improve the ability that its energy longer-term bears field conditions.

The compound for increasing the level of Sirtuin and/or the regulation Sirtuin of activity can also be used for increase Plant life, stress resistance and the resistance for Apoptosis.In one embodiment, compound is bestowed into plant (for example Periodically), or fungi is bestowed.In other embodiments, plant is genetically modified to produce compound.In another implementation In scheme, plant is handled in harvesting with fruit with first before transport with compound, to increase during transporting for the anti-of damage Property.Also vegetable seeds can be contacted with compound as described herein, for example to enable it to preserve.

In other embodiments, the regulation Sirtuin of the level of increase Sirtuin and/or activity Compound can be used for the life-span of regulation yeast cells.Wherein it is desirable to the situation in extension yeast cells life-span includes wherein using yeast Any technical process, the preparation of such as beer, Yoghourt and baked items (such as bread).Using with prolonging long-life ferment Mother, can use less yeast, or the time for making yeast active is longer.Also can by for be prepared by recombinant albumen yeast or Other mammalian cells are handled as described herein.

The compound for increasing the level of Sirtuin and/or the regulation Sirtuin of activity can also be used for increase The insect life-span, stress resistance and the resistance for Apoptosis.In this embodiment, compound will be administered to useful elder brother Worm, such as honeybee and other insects for being related to plant pollination.In a specific embodiment, compound will be administered to and relate to And the honeybee of production honey.In general, method described herein and purposes can be applied to any organism, such as with business The eucaryote of importance.For example, compound as described herein can be applied to fish (aquaculture) and birds (such as chicken with Birds).

The level of the increase Sirtuin of higher dosage and/or the regulation Sirtuin of activity can also be used Compound is as insecticide, and this is carried out by the regulation of cryptiogene during disturbing development and the regulation of Apoptosis. In this embodiment, the compound can be applied to plant by methods known in the art, and ensures the compound It is biological available for insect larvae (and not being for plant).

At least for the viewpoint contacted between reproduction and life-span, can apply increase Sirtuin level and/or The compound of the regulation Sirtuin of activity, to influence the reproduction of the organisms such as insect, animal and microorganism.

Other embodiments

In one aspect, the present invention relates to the method for the activity of increase Sirtuin -1 in cell, including cell is made Respectively with formula (I) compound or its pharmaceutically acceptable salt or its corresponding pharmaceutical composition thereof the step of.

In one aspect, the present invention relates to treatment insulin resistance, metabolic syndrome, diabetes or their complication or Increase the method for insulin sensitivity, including give compound respectively to subject in need or its is pharmaceutically acceptable Salt or its corresponding pharmaceutical composition.

In one aspect, the present invention relates to the method for the treatment of metabolic dysfunction, including to subject in need point Compound or its pharmaceutically acceptable salt or its corresponding pharmaceutical composition are not given.

In one aspect, expressed the present invention relates to treatment by SIRT1 or activity reduce caused disease or the method for obstacle, It includes giving compound or its pharmaceutically acceptable salt or its corresponding medicine respectively to subject in need Composition.

In one aspect, the present invention relates to a kind of method, wherein reducing caused disease or barrier by SIRT1 expression or activity Hinder and be selected from, but not limited to, aging or stress, diabetes, metabolic dysfunction, neurodegenerative disease, angiocardiopathy, cancer or Inflammatory disease.

In one aspect, the present invention relates to a kind of method, wherein with aging or stress be relevant disease, diabetes, metabolism Dysfunction, neurodegenerative disease, angiocardiopathy, cancer or inflammatory disease are selected from psoriasis, atopic dermatitis, acne, wine Slag nose, inflammatory bowel disease, osteoporosis, septicemia, arthritis, COPD, systemic loupus erythematosus and ophthalmic inflammation.

In one aspect, the present invention relates to kind of a method, wherein with aging or stress be relevant disease, diabetes, metabolism work( Energy obstacle, neurodegenerative disease, angiocardiopathy, cancer or inflammatory disease are selected from psoriasis, atopic dermatitis, acne, schlempe Nose, inflammatory bowel disease, osteoporosis, septicemia, arthritis, COPD, systemic loupus erythematosus and ophthalmic inflammation.

In one aspect, the present invention relates to treatment psoriasis method, it include to subject in need respectively to Give compound or its pharmaceutically acceptable salt or its corresponding pharmaceutical composition.

In one aspect, the present invention relates to give compound or its pharmaceutically acceptable salt or its corresponding medicine respectively Composition, in therapy, insulin resistance, metabolic syndrome, diabetes are suffered from or are susceptible to suffer from or theirs is concurrent for treating The subject of disease or for increasing insulin sensitivity in subject.

In one aspect, the present invention relates to give compound or its pharmaceutically acceptable salt or its corresponding medicine respectively Composition is being prepared for treating insulin resistance, metabolic syndrome, diabetes or their complication or in subject Purposes in the medicine of middle increase insulin sensitivity.

Determine

The other methods and purposes covered herein include be used for identify regulation Sirtuin compound or The screening technique of medicine.Reagent can be nucleic acid, such as fit (aptamer).Measure can be based on cell or not celliferous shape Formula is carried out.The bar that Sirtuin can be adjusted by the reagent place of known regulation Sirtuin is can be included in for example, determining Under part, Sirtuin is incubated (or contact) with test agent, and relative to test agent is lacked, deposited in the test agent In lower monitoring or the regulation level of measure Sirtuin.The regulation level of Sirtuin can be by determining it by substrate Deacetylated ability and determine.Example substrate is the acetylation for being purchased from BIOMOL (Plymouth Meeting, PA) Peptides.Preferred substrate includes p53 peptides, for example those peptides for including acetylation K382.Especially preferred substrate is Fluor De Lys-SIRT1 (BIOMOL), i.e. acetylated peptide Arg-His-Lys-Lys.Other substrates are derived from human histone H3 and H4 Peptides or acetylated amino acids.Substrate can be (fluorogenic) of fluorescence.The Sirtuin can for SIRT1, Sir2, SIRT3 or part thereof.For example, restructuring SIRT1 is available from BIOMOL.Reaction can be carried out about 30 minutes, and for example with nicotinoyl Amine is terminated.Can be used HDAC fluorescence activities measure/drug discovery kit (drug discovery kit) (AK-500, BIOMOL Research Laboratories) determine Acetylation Level.Similar measure is described in Bitterman et al. (2002)J.Biol.Chem.277：In 45099.Can be by the regulation level of the Sirtuin in measure and a kind of or many Kind (separately or simultaneously) Sirtuin in the presence of compound (it can be used as positive or negative control group) described herein Regulation level is compared.Sirtuin for measure can be total length Sirtuin or part thereof.Because at this Show that activating compounds show the N- end effects with SIRT1 in text, therefore adjusted for the albumen of the measure including silence Save the N- end sections of albumen, such as SIRT1 substantially amino acid/11-176 or 1-255；Sir2 substantially amino acid/11- 174 or 1-252 parts.

In one embodiment, screening test is being adapted to the Sirtuin in the presence of without test agent including (i) Under conditions of substrate deactylation, Sirtuin is set to be contacted with test agent with acetylation substrate；And (ii) determines bottom The Acetylation Level of thing, wherein in the presence of the degree of acetylation of substrate is relative to without the test agent in the presence of the test agent It is relatively low, represent：The test agent stimulates the deacetylated effect carried out by Sirtuin, and exists in the test agent The degree of acetylation of lower substrate is represented relative to without higher in the presence of the test agent：The test agent suppresses to be adjusted by silence The deacetylated effect that albumen is carried out.

In another embodiment, the NAD dependences of the detectable Sirtuin mediation of the screening test are deacetylated 2 '/3 '-O- acetyl group-ADP- ribose products formation.O- acetyl group-ADP- the ribose products are with de- with Sirtuin The deacetylated peptide prod equimolar amounts of acetylization reaction is formed.Correspondingly, the screening test may include that (i) is being adapted to without survey Sirtuin in the presence of agent of having a try, by under conditions of substrate deactylation, makes Sirtuin and test agent and acetyl Change substrate contact；And (ii) determines the amount of O- acetyl group-ADP- ribose formation, wherein O- acetyl group-ADP- ribose formation is being surveyed Amount increase during relative in the absence of the test agent of having a try in the presence of agent is then represented：The test agent stimulates and adjusts egg by silence The deacetylated effect carried out in vain, and O- acetyl group-ADP- ribose is formed in the presence of test agent relative in the absence of the survey Have a try agent when amount reduction then represent：The test agent suppresses the deacetylated effect carried out by Sirtuin.

The method and purposes of the reagent of Sirtuin (is for example stimulated) to may include that (i) exists in internal regulation for identifying In the presence of I classes and II classes HDAC inhibitor, suitable Sirtuin in the presence of without test agent substrate is deacetylated Under conditions of change, make cell and test agent and the substrate of cell can be entered to contact；And (ii) determines the acetylation water of substrate It is flat, wherein it is relatively low in the presence of the Acetylation Level of substrate is relative to without the test agent in the presence of the test agent, represent： The test agent stimulates the deacetylated effect that is carried out by Sirtuin, and in the presence of the test agent substrate acetyl Change horizontally relative to without higher in the presence of the test agent, represent：The test agent suppresses by taking off that Sirtuin is carried out Acetylation.Preferred substrate is acetylated peptide, and it is preferably also fluorescence, as further described herein.This method can Further comprising cell is cracked into (lysing), to determine the degree of acetylation of substrate.Can by substrate with scope between about 1 μM extremely About 10mM, preferably about 10 μM to 1mM, even more preferably about 100 μM to 1mM, e.g., from about 200 μM of concentration is added in cell. Preferred substrate is acetylated lysine, for example ε-acetyllysine (Fluor de Lys, FdL) or Fluor de Lys- SIRT1.I classes and II classes HDAC preferred inhibitor are Trichostatin A (trichostatin A) (TSA), and it can be with scope Between about 0.01 μM to 100 μM, preferably from about 0.1 μM to 10 μM, such as 1 μM of concentration is used.Cell and test compound and bottom The incubation of thing can carry out about 10 minutes to 5 hours, preferably from about 1-3 hours.Because TSA suppresses all I classes and II class HDAC, and one A little substrate such as Fluor de Lys are poor substrate for SIRT2, are even worse substrate for SIRT3-7, so Class determines the conditioning agent that can be used for identifying internal SIRT1.

Method and purposes in therapy

The invention further relates to use Sirtuin conditioning agent compounds for treating as defined herein and/or the wide model of prevention The a variety of diseases and the method and purposes of obstacle enclosed, the disease and obstacle include but is not limited to, for example, with aging or stress be anti- Disease or obstacle that should be relevant, diabetes, obesity, neurodegenerative disease, angiocardiopathy, blood coagulation disorder, inflammation, cancer Disease and/or flush, and mitochondrial active increased disease or obstacle are benefited from, the disease and obstacle are further selected from or wrapped Include but be not limited to psoriasis, atopic dermatitis, acne, brandy nose, inflammatory bowel disease, osteoporosis, septicemia, arthritis, COPD, systemic loupus erythematosus and ophthalmic inflammation.

On the other hand, adjusted the invention provides the compound using regulation Sirtuin or comprising regulation silence The method and purposes of the composition of the compound of albumen.In certain embodiments, improve Sirtuin level and/or The compound of the regulation Sirtuin of activity can be used for various treatment uses, including, for example, increase cell survival, is controlled The a variety of diseases and illness of wide scope are treated and/or prevent, including, for example, the disease relevant with aging or stress reaction or obstacle, Diabetes, obesity, neurodegenerative disease, the neuropathy of chemotherapy-induced, the neuropathy relevant with ischemic event, eyes disease Disease and/or illness, angiocardiopathy, blood clotting illness, inflammation, and/or flush, etc..Improve the water of Sirtuin The mitochondria activity of benefiting from that the compound of flat and/or activity regulation Sirtuin can be used for treating patient increases Disease or obstacle, for strengthening muscle performance, for improving muscle ATP levels, or for treating or preventing and anoxic or lacking The relevant muscle tissue damage of blood.In other embodiments, the regulation of the level and/or activity of reduction Sirtuin is sunk The compound of silent regulatory protein can be used for various treatment uses, including, for example, sensitiveness of the cell to stress reaction is improved, Increase Apoptosis, treating cancer stimulates appetite, and/or stimulate increased weight, etc..As described further below, it is described Method and purposes include the compound that the regulation Sirtuin of pharmaceutical effective amount is administered to subject in need.

In some respects, it is described regulation Sirtuin compound can be administered alone combined with other compounds to Medicine, other described compounds include the compound or other therapeutic agents of other regulation Sirtuins.

On the other hand, the present invention relates to the method for the activity of Sirtuin -1 in increase cell, it includes making cell The compound of the formula (I) to (IV) of the present invention is contacted, its corresponding analog or derivative are (that is, in R²Position has hydrogen substitution Analog) the step of.

On the other hand, the present invention relates to the method for the activity of Sirtuin -1 in increase cell, including connect cell The step of touching pharmaceutical composition of the invention defined herein.

On the other hand, the present invention relates to treatment insulin resistance, metabolic syndrome, diabetes or their complication or Increase the method for insulin sensitivity, it includes the chemical combination that the formula (I) to (IV) of the present invention is given to subject in need Thing, its corresponding analog or derivative are (that is, in R²Position has the analog that hydrogen replaces) the step of.

On the other hand, suffer from or be susceptible to suffer from the present invention relates to treatment insulin resistance, metabolic syndrome, diabetes or they Complication subject or in subject increase insulin sensitivity method, including by the present invention drug regimen Thing gives subject in need

On the other hand, the present invention relates to treatment insulin resistance, metabolic syndrome, diabetes or their complication or Increase the method for insulin sensitivity, including subject in need is given by the pharmaceutical composition of the present invention.

On the other hand, the present invention relates to the method for the activity of Sirtuin -1 in increase cell, it includes making cell The compound of the formula (I) to (IV) of the present invention is contacted, its corresponding analog or derivative are (that is, in R²Position has hydrogen substitution Analog) or the step of its pharmaceutically acceptable salt.

On the other hand, the present invention relates to the method for the activity of Sirtuin -1 in increase cell, it includes making cell The step of contacting the pharmaceutical composition of the present invention.

On the other hand, the present invention relates to the method for the treatment of metabolic dysfunction, it is included to subject in need Formula (I) is given to the compound of (IV), its corresponding analog or derivative (that is, in R²Position has the analog that hydrogen replaces) Or the step of its pharmaceutically acceptable salt.

On the other hand, the present invention relates to treatment metabolic dysfunction method, including by the present invention pharmaceutical composition Give subject in need.

On the other hand, expressed the present invention relates to treatment by SIRT1 or activity reduce caused disease or the method for obstacle, It includes giving formula (I) to the compound of (IV) to subject in need, and its corresponding analog or derivative (that is, exist R²Position has the analog that hydrogen replaces) or its pharmaceutically acceptable salt.

On the other hand, the present invention relates to a kind of method, wherein described reduce caused disease by SIRT1 expression or activity Or obstacle is selected from, but not limited to, aging or stress, diabetes, metabolic dysfunction, neurodegenerative disease, angiocardiopathy, cancer Disease or inflammatory disease.

On the other hand, the present invention relates to a kind of method, wherein it is described with aging or stress be relevant disease, diabetes, Metabolic dysfunction, neurodegenerative disease, angiocardiopathy, cancer or inflammatory disease are selected from psoriasis, atopic dermatitis, Cuo Sore, brandy nose, inflammatory bowel disease, osteoporosis, septicemia, arthritis, COPD, systemic loupus erythematosus and ophthalmic inflammation.

On the other hand, the present invention relates to the method for the treatment of psoriasis, it is included to subject in need's giving construction The compound of (I) to (V), its corresponding analog or derivative are (that is, in R²Position has the analog that hydrogen replaces) or its medicine Acceptable salt on.

On the other hand, the present invention relates to the method for the treatment of psoriasis, it includes giving this to subject in need The pharmaceutical composition of invention.

Pharmaceutical composition and preparation

Generally, the present invention relates to the substituted bridging urea analog compounds of formula (I) to (V), its corresponding analog or Derivative is (that is, in R²Position has the analog that hydrogen replaces), or its pharmaceutically acceptable salt, corresponding pharmaceutical composition, The method for preparing the compound, and the compound is independent or is adjusted with combination with other therapeutic agents as Sirtuin Agent is used for following purposes：Improve cell survival, and treatment and/or a variety of diseases and obstacle of preventing wide scope, the disease Disease and obstacle include but is not limited to, for example, the disease relevant with aging or stress reaction or obstacle, diabetes, obesity, nerve Degenerative disease, angiocardiopathy, blood coagulation disorder, inflammation, cancer and/or flush, and benefit from mitochondrial activity increasing Plus disease or obstacle.

In particular it relates to the noval chemical compound of formula (I) to (V), its corresponding analog or derivative are (that is, in R² Position has the analog that hydrogen replaces) or its pharmaceutically acceptable salt, and the phase of formula (I) to the compound of (V) is included respectively The pharmaceutical composition answered.

On the other hand, the present invention relates to pharmaceutical composition, it includes pharmaceutically acceptable carrier or diluent and formula The compound of (I) to (V), its corresponding analog or derivative are (that is, in R²Position has the analog that hydrogen replaces) or its medicine Acceptable salt on.

On the other hand, the present invention relates to the pharmaceutical composition of the present invention, it also includes other activating agents.

On the other hand, the present invention relates to pharmaceutical composition, the compound of its bag formula (I) to (V), its corresponding analog Or derivative is (that is, in R²Position have hydrogen replace analog) or its pharmaceutically acceptable salt and at least one pharmaceutically may be used The carrier of receiving.

Compound as described herein can be physiologically or pharmaceutically acceptable using one or more by conventional method Carrier or excipient.For example, compound and its pharmaceutically acceptable salt can be configured to be used to pass through with solvate (such as SubQ, IM, IP) is for example injected, sucks or is blown into (by mouth or nose) or oral, buccal, sublingual, percutaneous, nose, stomach The preparation of outer or rectally.In some embodiments, the compound can partly, the position existed in target cells, It is administered in particular organization, organ or body fluid (such as blood, celiolymph).

Compound can be configured to be used for various administering modes, include the preparation of whole body and part or regional administration.Technology Remington ' s Pharmaceutical Sciences, Meade Publishing Co. are generally found in preparation, Easton, PA.Be preferred, including intramuscular, intravenous, intraperitoneal and subcutaneously to inject for parenteral.For note Penetrate, the compound can be configured to liquid solution, preferably with physiologically compatible buffer solution, such as Hank's solution or woods Grignard solution.The compound can be configured to solid form in addition, and be redissolved or suspend immediately in before.It may also comprise Freeze-dried.

For being administered orally, pharmaceutical composition can be used for example by conventional method, use pharmaceutically acceptable excipient Such as adhesive (for example, the cornstarch of pregelatinized, PVP or hydroxypropyl methyl cellulose)；Filler (such as lactose, microcrystalline cellulose or calcium monohydrogen phosphate)；Lubricant (such as magnesium stearate, talcum or silica)；Disintegrant (such as horse Bell sweet potato starch or sodium starch glycolate)；Or wetting agent (such as NaLS) prepare tablet, the shape of lozenge or capsule Formula.Tablet can be coated by method known to this area.Liquid preparation for oral administration can use such as solution, syrup Or the form of suspension, or they can be made into desciccate, be constructed using preceding with water or other appropriate carriers.This class I liquid I system Agent can be by conventional method, with pharmaceutically acceptable additive such as suspending agent (for example, sorbitol syrup, cellulose spread out Biological or hydrogenated edible fats class)；Emulsifying agent (such as lecithin or Arabic gum)；Nonaqueous intermedium (such as almond oil, ester oil Class, ethanol or classification vegetable oil)；And preservative (such as p-hydroxy Benzoic Acid methyl ester or p-hydroxy Benzoic Acid ester propyl diester Or sorbic acid) prepare.If appropriate, said preparation can also contain buffer salt, flavor enhancement, colouring agent and sweetener.For orally giving The preparation of medicine can be prepared suitably, so that reactive compound controllably discharges.

Suck (such as lung delivering) for passing through and be administered, it is convenient to by compound in aerosol spray presentation form from warp There is provided and deliver in pressurized package or sprayer, wherein using suitable propellant for example dicholorodifluoromethane, Arcton 11, Dichlorotetra-fluoroethane, carbon dioxide or other suitable gases.In the case of pressurized aerosol, dosage unit can be by providing Determined for delivering the valve for the amount measured.Such as gelatine capsule or cartridge case of inhalator or insufflator can be formulated for (cartridge), its mixture of powders for having compound and suitable powder base such as lactose or starch containing described in.

Compound can be formulated into for by injection (such as by inject or continuous infusion) parenteral.For noting The preparaton penetrated can be provided in a unit, for example, being provided with ampoule or multi-dose container (addition preservative).Composition Can be using the suspension such as in oiliness or aqueous vehicle, the form of solution or emulsion, and preparaton can be contained, for example hang Floating agent, stabilizer and/or dispersant.Or, active component can be powder type, and convenient medium thing can be used before use, For example sterile non-pyrogenic water is constructed.

The compound can also be configured to rectal compositions, such as suppository or enema,retention, such as containing conventional bolt Agent matrix such as cocoa butter or other glyceride types.

In addition to previously described preparation, compound can also be configured to depot formulations (depot preparation).It is such Depot formulations can be by implantation (such as with subcutaneous or intramuscular), or via intramuscular administration.Thus, for example, will can change Compound is prepared with suitable polymerization or hydrophobic material (such as the emulsion in acceptable oil) or ion exchange resin, or in micro- Soluble derivatives form, such as in slightly soluble salt form.Controlled release formulation also includes patch.

In some embodiments, compound as described herein can be configured to for delivery to central nervous system (CNS) (see summary, Begley, Pharmacology＆Therapeutics 104：29-45(2004)).For medicine delivery to CNS's Conventional method includes：Neurosurgery strategy (such as intracerebral injection or intraventricular delivery)；Molecule manipulation (such as generation bag of reagent Containing the chimeric fusion protein with the transit peptides of pharmaceutical agent combinations, the transit peptides Human Umbilical Vein Endothelial Cells surface molecular has affinity, and The medicament can not pass through BBB in itself), one of endogenous transport approach to attempt exploitation BBB；Designed for increase reagent The pharmacology strategy (for example, (conjugation) is conjugated to lipid or cholesterol carrier in water-soluble reagent) of liposolubility；And pass through The hypertonic of short duration destruction BBB integralities of destruction (are derived from and mannitol solution are infused into arteria carotis or using bioactivator example Such as angiotensin).

Liposome is the drug delivery system of another easy injectable.Then, in the methods of the invention also can be by activity Compound is administered in the form of liposome delivery system.Liposome is known to those skilled in the art.Liposome can Formed by the stearylamine of various phosphatide such as cholesterol, phosphatldylcholine class.Liposome available for the inventive method includes institute There is type liposome, including but not limited to small monolayer vesicle, big monolayer vesicle and multi-layer vesicles.

The method of another preparation (especially solution) for preparing compound as described herein is by using cyclodextrin.Ring Dextrin refer to α-, β-or gamma-cyclodextrin.Cyclodextrin is described in detail in Pitha et al., U.S patents 4,727,064.Cyclodextrin For the cyclic oligomeric thing of glucose；These compounds and any medicine formation inclusion complex (inclusion complex), institute Stating the molecule of medicine can adapt in the lipophilic search cavity (lipophile-seeking cavity) of cyclodextrin molecular.

Fater disintegration or dissolving formulation can be used for the quick absorption of pharmaceutical active, especially buccal and sublingual absorption.It hurry up Instant solution (fast melt) formulation is to the patient difficult with tablet with common solid dosage forms such as caplet (caplet) is swallowed (such as the elderly and young children) is beneficial.In addition, rapid-dissolve dosage form overcomes the shortcoming relevant with such as chewable dosage forms, its The time span that middle activating agent is kept in the patient's mouth is it is determined that the sense of taste amount of covering and patient may experience the throat of activating agent Played an important role in terms of the degree of coarse sand texture.

Pharmaceutical composition (including cosmetic formulations) can include about 0.00001% to 100%, such as 0.001 to 10% or One or more compounds as described herein of 0.1% to 5% (by weight).In other embodiments, the drug regimen Thing is included：(i) 0.05 to 1000mg compound of the invention or its pharmaceutically acceptable salt, and 0.1 to 2 gram of (ii) one Plant or a variety of pharmaceutically acceptable excipient.

In some embodiments, compound as described herein is mixed into containing the office for applying in general to topical drug administration In portion's drug administration carrier and topical formulations comprising any such material known in the art.Part drug administration carrier may be selected so that group Compound is in desired form, and such as in ointment, lotion, emulsifiable paste, microemulsion, gel, oil, solution, and it can be by naturally depositing Or synthesis source material composition.It is preferred that selected carrier is not adversely affected by other groups of activating agent or topical formulations Point.For this paper suitable local drug administration carrier example include water, alcohols and other non-toxic organic solvents, glycerine, mineral oil, Silicone, vaseline, lanolin, aliphatic acid, vegetable oil, p-hydroxybenzoate, wax class etc..

Preparation can be ointment, lotion, emulsifiable paste, microemulsion and the gel of colorless and odorless.

The compound can be mixed into the generally ointment of semisolid preparation, and it is generally with vaseline or other petroleum derivations Thing is matrix.The specific ointment bases that used those skilled in the art are understood is can to provide optimal drug delivering, and It is preferred that the matrix of other desired features and such as property of softening or similar characteristics can be provided.With other carriers or excipient one Sample, ointment bases should be inertia, stably, nonirritant and non-sensitization.

The compound can be mixed into lotion, and it is generally will be applied to the preparation of skin surface without friction, and be usually Wherein solid particle (including activating agent) is present in liquid or semi liquid state preparation in water or alcohol base.Lotion is normally solid Suspension, and may include the liquid oily emulsion of oil-in-water type.

The compound can be mixed into emulsifiable paste, and it is generally liquid of vicidity or semisolid emulsion, be oil-in-water or oil bag Water type.Emulsion bases is washable, and contains oil phase, emulsifying agent and aqueous phase.Oil phase it is general by vaseline and fatty alcohol (for example Cetanol or stearyl alcohol) constituted；Aqueous phase generally (although not necessarily) exceedes oil phase in volume, and typically contains wetting agent. Emulsifying agent in cream preparation is as illustrated in foregoing Reminton document, generally non-ionic, anionic, sun Ionic or amphoteric surfactant.

The compound can be mixed into microemulsion, and it is generally two kinds of not miscible liquid (such as oil and water) via table The stable, dispersion of isotropic clarification in the stabilized thermokinetics of interfacial film of face active agent molecule (Encyclopedia of Pharmaceutical Technology(New York：Marcel Dekker, 1992), the 9th Volume).

The compound can be mixed into gel preparation, and it is generally semi-solid systems, and the semi-solid systems are by small inorganic Molecular suspension (two-phase system) composition of grain, or by being substantially uniformly distributed in whole carrier liquid (single-phase gels) Big organic molecule is constituted.Although gel typically uses aqueous carrier liquid, it is possible to use alcohols is used as carrier fluid with oils Body.

May also comprise other activating agents in the formulation, such as other antiphlogistics, analgestic, antimicrobial, antifungal agent, Antibiotic, vitamin, antioxidant and the sun-block agent being generally present in sun-screening agent, including but not limited to adjacent amino Benzoic ether, benzophenone (especially BP-3), camphor derivatives, (for example methoxycinnamate is misery for cinnamate Ester), dibenzoyl methane (such as butylmethoxydibenzoylmethane), Para-Aminobenzoic (PABA) and its derivative Thing, and salicylate (such as octyl salicylate).

In some topical formulations, activating agent is with about 0.25 weight % of said preparation to 75 weight %, it is therefore preferable to should About 0.25 weight % of preparation to 30 weight %, more preferably said preparation about 0.5 weight % are to 15 weight %, and most Preferably about 1.0 weight % of said preparation to 10 weight % scopes amount exist.

Eye diseases can be for example, by systemic, part, intraocular injection compound, or holding by embedded releasable compound Continue release device and treat or prevent.It will can be delivered in compound pharmaceutically acceptable eye medium, so that described Compound can keep contacting time enough to allow the compound to penetrate the interior zone of cornea and eyes, for example with eye surface Cup, rear chamber, vitreum, aqueous humor, vitreous humor, cornea, iris/ciliary body (ciliary), crystalline lens, choroid/retina And sclera.Pharmaceutically acceptable eye medium may be, for example, ointment, vegetable oil or encapsulating material.On the other hand, this can be sent out Bright compound is injected directly into vitreous humor and aqueous humor.In another selection, the compound can systemic applications (for example lead to Cross intravenous infusion or injection) it is used for the treatment of eye.

Compound as described herein can be stored in oxygen-free environment.For example, composition can be formulated in for oral administration In seal capsule, such as purchased from Pfizer, Inc. Capsugel.

Can be by the cell of use-case Compound ira vitro processing as described herein according to for graft to be delivered medicine into patient Method be administered, its can with for example be administered immunosuppressive drug such as cyclosporin A.The general original prepared on medicine Reason, reader refers to Cell Therapy：Stem Cell Transplantation, Gene Therapyand Cellular Immunotherapy, G.Morstyn and W.Sheridan are compiled, Cambridge University Press, and 1996；And Hematopoietic Stem Cell Therapy, E.D.Ball, J.Lister and P.Law, Churchill Livingstone, 2000.

The toxicity of compound can be surveyed with therapeutic efficiency by with standard pharmaceutical procedures in cell culture or experimental animal It is fixed.LD₅₀To make the dosage that 50% colony is lethal.ED₅₀For for effective dosage in 50% mass treatment.Toxicity and treatment work( Dose ratio (LD50/ED50) between effect is therapeutic index.It is preferred that the compound with high therapeutic index.Although can be used has The compound of toxic side effect, but careful design is answered by the delivery system of such targeting compounds to infected tissue position, so that Obtaining can will reduce to minimum for the possible injury of uninfection cell, and thus reduce side effect.

Data derived from cell culture test and zooscopy can be used for preparing the dosage range used in people.Suchization The dosage of compound can include having very small toxicity or avirulent ED at it₅₀In the range of the circulation composition of value.The dosage can be It is varied from the range of this according to used formulation and the method for administration utilized.For any compound, can initially by Cell culture test assesses the upper effective dosage for the treatment of.Animal model allocating dosage can be used, to reach including such as in cell culture Middle determined IC₅₀The circulating plasma of value (that is, the test compound concentration for reaching the half maximum suppression effect to symptom) is dense Spend scope.This type of information can be used for more accurately determining the effective dose to people.For example it can be measured by high performance liquid chromatography Concentration in blood plasma.

Kit

The present invention also provides kit, for example kit, or for therapeutic purposes thin for adjusting cell survival or regulation The kit of born of the same parents' apoptosis.Kit may include one or more for example with the chemical combination specifically described herein of the dosage measured in advance Thing.Kit optionally includes the device and operation instructions for being used to contact cell with the compound.The device includes Syringe, support and it is other be used for compound is incorporated into patient's (such as blood vessel of patient) or is coated on patient skin Device.

In another embodiment, the present invention provides a kind of composition, its compound comprising the present invention with it is another Therapeutic agent (with for combination treatment and combine those identical therapeutic agents in composition), they are present in independent formulation But associate each other.Terms used herein " interrelated " refers to, independent formulation is packaged together, or mutually according to It is attached, so that being readily apparent that these independent formulations are intended to be sold and be intended to the part administration as same approach. It is preferred that the compound and other medicaments are packaged in blister package or other multicells packaging together, or conduct can be by user Voluntarily separate link, separated sealed container (such as tinfoil paper of (for example, by being torn at the score line between two containers) Pouch etc.).

In another embodiment, the present invention provides kit, and it includes change a) of the invention in independent container Compound；And b) another therapeutic agent, such as those therapeutic agents for being described in specification other parts.

Unless otherwise indicated, the implementation of the inventive method will utilize the cell biological known to those of ordinary skill in the art , cell culture, molecular biology, transgcnic biology, microbiology, recombinant DNA and immunologic routine techniques.These skills Art has detailed explanation in the literature.See, e.g.：Molecular Cloning ALaboratorymanual, second edition, Sambrook, Fritsch and Maniatis write (Cold Sp Spring Harbor Laboratory Press：1989)； DNA Cloning, I and II volumes (D.N.Glover writes, 1985)；Oligonucleotide Synthesis(M.J.Gait Write, 1984)；Mullis et al. United States Patent (USP)s：4,683,195；Nucleic Acid Hybridization(B.D.Hames& 1984) S.J.Higgins writes；Transcription And Translation (B.D.Hames＆S.J.Higgins writes, 1984)；Culture of Animal Cells (R.I.Freshney, Alan R.Liss, Inc., 1987)；Immobilized Cells And Enzymes (IRLPress, 1986)；B.Perbal, Practical Guide To Molecular Cloning(1984)；Disquisition, Methods In Enzymology (Academic Press, Inc., N.Y.)；Gene Transfer Vectors For Mammalian Cells (J.H.Millerh and M.P.Calos write, and 1987, Cold Spring Harbor Laboratory)；Methods In Enzymology, roll up 154 and 155 (Wu et al. writes), Immunochemical Methods In Cell And Molecular Biology (Mayer and Walker write, Academic Press, London, 1987)；Handbook of Experimental Immunology, roll up I-IV (D.M.Weir and C.C.Blackwell write, 1986)；Manipulating the Mouse Embryo(Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1986).

Embodiments illustrated below is the description of the invention, the scope being not intended in any way to limit the present invention.

Embodiment

Embodiments illustrated below is the description of the invention, the scope being not intended in any way to limit the present invention, But the compound of the present invention is prepared and used for those skilled in the art, composition and method or purposes provide guidance.

Although describing the particular embodiment of the present invention, it will be appreciated, however, by one skilled in the art that not departing from the present invention Spirit and scope in the case of, can make various changes and modifications.

Such as this paper symbols used in methods described, scheme and embodiment and convention and contemporary scientific literature (for example Journal of the American Chemical Society or the Journal of Biological Chemistry the symbol used in) is consistent with convention.Amino is represented usually using Standard single-letter or three alphabetical abbreviations Sour residue, unless otherwise stated, it is L- configurations that these abbreviations, which are assumed,.Unless otherwise indicated, all initiation materials by Commercial supplier is obtained, and can be used without further purification.

It is all that refer to ether is ether；Salt solution refers to NaCl saturated aqueous solution.Unless otherwise indicated, all temperature with DEG C (degree Celsius) represent.Unless otherwise stated, all reactions are carried out at room temperature under an inert atmosphere, unless otherwise saying Bright, all solvents are available highest purity.

The equipment used

LCMS with PDA:

Waters Allaince2695-2996/Quattromicro

Agilent-1200/SQD

Preparative LC (preparation HPLC) with UV detectors:

Waters-2545/2998PDA and 2487UV

Shimadzu–LC-20AP/20AV-UV

Gilson-333,334/115-UV

Chiral HPLC:

Waters Alliance-2695/2998&2996

SFC purification systems:

Thar-SFC-80

Waters SFC–200

NMR(400MHz):

Varian-400MHz

Use 2014ACD laboratories Software Create¹H-NMR schemes.

Recorded on Varian-400MHz spectrometers¹H NMR (hereinafter also referred to " NMR ") spectrum.Chemical shift is with million / mono- (ppm, δ unit) is represented.Coupling constant unit is hertz (Hz).Split merotype and show apparent multiplicity, and referred to It is set to s (singlet), d (doublet), t (triplet), q (quartet), quint (quintet), m (multiplet), br (broad peak).

The LCMS methods used

Acq. method condition:RND-ABC-6-MIN.

Post：XBridge BEH C18(50mm×4.6mm,2.5μm)

Mobile phase:A:5mM ammonium bicarbonate aqueous solutions (PH-10 contains ammoniacal liquor):ACN

Time (min)/%ACN:0/5,0.5/5,1/15,3.3/98,5.2/98,5.5/5,6.0/5

Column temperature：35 DEG C, flow velocity 1.3ml/min.

MS parameters:

Mass range:100-1000

Sweep time:0.5 second

Scanning room postpones:0.1 second

Run time:6.0min.

Acq. method condition:RND-FA-4.5-MIN.

Post：Acquity BEH C18(50mm×2.1mm,1.7um)

Mobile phase:A:The 0.1%FA aqueous solution；B:0.1%FA/ACN

Time (min)/%B:0/3,0.4/3,3.2/98,3.8/98,4.2/3,4.5/3

Column temperature：35 DEG C, flow velocity:0.6mL/min.

MS parameters:

Mass range:100-1000

Sweep time:0.5 second

Scanning room postpones:0.1 second

Run time:4.5min.

Acq. method condition:RND-FA-4.5-MIN.

Post：Acquity BEH C18(50mm×2.1mm,1.7um)

Mobile phase:A:The 0.1%FA aqueous solution；B:0.1%FA/ACN

Time (min)/%B:0/3,0.4/3,3.2/98,3.8/98,4.2/3,4.5/3

Column temperature：35 DEG C, flow velocity:0.6mL/min.

MS parameters:

Mass range:100-1000

Fragmentation voltage (Fragmentor):100

Step-length:0.1

Run time:4.5min.

Acq. method condition:RND-ABC-6.5-MIN.

Post：XBridge BEH C18(50mm×4.6mm,2.5μm)

Time (min)/%ACN:0/5,0.5/5,1/15,3.3/98,6.0/98,6.1/5,6.5/5

Column temperature：35 DEG C, flow velocity 1.3ml/min

MS parameters:

Mass range:100-1000

Fragmentation voltage:100

Step-length:0.1

Run time:6.5min.

Acq. method condition:RND-ABC-10-MIN.

Post：XBridge BEH C18(50mm×4.6mm,2.5μm)

Time (min)/%ACN:0/5,0.5/5,1.5/15,7/98,9.0/98,9.5/5,10/5

Column temperature：35 DEG C, flow velocity 1.3ml/min.

MS parameters:

Mass range:100-1000

Fragmentation voltage:100

Step-length:0.1

Run time:10.0min.

Intermediate

Synthesize bicyclic pyridine core

Synthesize (S) -2- ((the chloro- 3- nitropyridines -2- bases of 6-) amino) dimethyl succinate

The chloro- 3- nitros pyrroles of 2,6- bis- are added into the 2L flasks equipped with thermometer, reflux condenser and mechanical stir Pyridine (100g, 0.52mol), (S)-aspartic acid diformazan ester hydrochloride (205g, 1.04mol), NaHCO₃(174g,2.07mol) With tetrahydrofuran (1L).Reactant mixture is stirred into 16h at 40 DEG C, the disappearance of 2,6- dichloropyridins is then monitored through HPLC. After the completion of reaction, solid is filtered out, (3 × 300mL) is then washed with ethyl acetate.The filtrate of merging and washings are concentrated into It is dry, residue is absorbed in 1L ethyl acetate.Solution is stirred into 2h together with charcoal (200g) in environment temperature, then by carbon filtration Remove, and washed with extra ethyl acetate (3 × 200mL).The filtrate of merging and washings are concentrated in vacuo, and acquisition thick material (S)- 2- ((the chloro- 3- nitropyridines -2- bases of 6-) amino) dimethyl succinate (180g,>100%), it is yellow oil.Its without Being further purified just is used for next step.LRMS(m/z):318.0[M+H]+；HRMS(m/z):[M+H]+C₁₁H₁₃N₃O₆Cl calculating Value, 318.0493；Measured value, 318.0492；¹H-NMR(300MHz,DMSO-d6):δ 9.00 (d, J=7.9Hz, 1H ,-NH), 8.50 (d, J=8.6Hz, 1H), 6.92 (d, J=8.6Hz, 1H), 5.23 (m, J=5.7,7.9Hz, 1H ,-CHNH), 3.67 (s, 3H), 3.63 (s, 3H), 3.06 (m, J=5.8Hz, 2H ,-CHCH₂)；¹³C-NMR(APT)(75MHz,DMSO-d6):δ170.93 (C),170.65(C),154.65(C),150.59(C),138.82(CH),127.28(C),112.81(CH),52.23(CH₃), 51.74(CH₃),50.20(CH),35.31(CH₂)。

Synthesize (S) -2- (6- chloro-2-oxo -1,2,3,4- tetrahydropyridines simultaneously [2,3-b] pyrazine -3- bases) acetoxymethyl ester

Load thick material (S) -2- into the 5L three-neck flasks equipped with thermometer, reflux condenser and mechanical stir ((the chloro- 3- nitropyridines -2- bases of 6-) amino) dimethyl succinate (180g, 0.52mol), iron powder (146g, 2.59mol), 2- Propyl alcohol (2L) and water (700mL).By mixture in 40 DEG C of stirrings, acetic acid (15.5g, 0.259mmol), adding speed are then added It is enough to keep internal temperature to be less than 70 DEG C.Reactant mixture is stirred into 30min at 70 DEG C, HPLC Indicator Reactions are completed.Will mixing Thing is cooled to 40 DEG C, then adds Na₂CO₃(165g, 1.55mol), stirs the mixture for 1h.Filter solid is crossed, then solid is used Tetrahydrofuran washs (3 × 500mL).The filtrate of merging and washings are concentrated in vacuo, and then by residue, (1L) is stirred in ethanol Mix 12 hours.Solid is filtered and washed with cold ethanol, is then dried in vacuo, acquisition (S) -2- (6- chloro-2-oxos -1,2, 3,4- tetrahydropyridines simultaneously [2,3-b] pyrazine -3- bases) acetoxymethyl ester, it is pale solid (91g, 68%).LRMS(m/z): 256.0[M+H]+；HRMS(m/z):[M+H]+C₁₀H₁₁N₃O₃Cl calculated value, 256.0489；Measured value, 256.0487；¹H- NMR(300MHz,DMSO-d6):δ 10.55 (br s, 1H ,-NHCO), 7.35 (br s, 1H ,-NHCH), 6.92 (d, J= 7.9Hz, 1H), 6.57 (d, J=7.8Hz, 1H), 4.43 (m, J=1.4,5.1Hz, 1H ,-NHCH), 3.57 (s, 3H ,- CO₂), Me 2.79 (m, J=5.1,16.4Hz, 2H ,-CHCH₂)；¹³C-NMR(APT)(75MHz,DMSO-d6):δ170.32(C), 164.96(C),146.13(C),140.32(C),122.41(CH),119.47(C),111.31(CH),51.81(CH),51.39 (CH₃),37.01(CH₂)。

Synthesize (S) -2- (the chloro- 1,2,3,4- tetrahydropyridines of 6- simultaneously [2,3-b] pyrazine -3- bases) ethanol

Load LiAlH into the 5L 3- neck flasks equipped with mechanical stir, reflux condenser and nitrogen inlet₄(60g, 1.58mol).Flask is cooled with an ice bath, and then adds tetrahydrofuran (500mL).The mixture of stirring is cooled to 0 DEG C, then Add (S) -2- (6- chloro-2-oxo -1,2,3,4- tetrahydropyridines simultaneously [2,3-b] pyrazine -3- bases) acetoxymethyl ester (81g, 0.32mol) the solution in tetrahydrofuran (2L), while keeping internal temperature to be less than 5 DEG C.After addition terminates, reaction is mixed Thing is heated at reflux 16h, while monitoring the appearance of product by HPLC.The rapid reduction for occurring ester, and lactam reduction needs more The long time completes.Reactant mixture is cooled to 5 DEG C, then water (60mL) is added while keeping internal temperature to be less than 10 ℃.After addition terminates, reactant mixture is stirred into 15min, 15% (w/w) NaOH (aqueous solution) (60mL) is then added, simultaneously Internal temperature is kept to be less than 5 DEG C.After addition terminates, reactant mixture is stirred into 15min, water (180mL) is then added, then will Mixture stirs 1h in environment temperature.Solid is filtered out, (3 × 150mL) is then washed with tetrahydrofuran.Filtrate and washings are true Solid residue, is then dried in vacuo by sky concentration, obtains (S) -2- (6- chloro- 1,2,3,4- tetrahydropyridines simultaneously [2,3-b] pyrrole Piperazine -3- bases) ethanol, it is brown solid (55g, 81%).LRMS(m/z):214.1[M+H]+；HRMS(m/z):[M+H]+ C₉H₁₃N₃OCl calculated value, 214.0747；Measured value, 214.0743；¹H-NMR(300MHz,DMSO-d6):δ6.60(br s, 1H,-NHCH(CH₂)₂), OH 6.58 (d, J=7.8Hz, 1H), 6.32 (d, J=7.8Hz, 1H), 5.69 (m, 1H ,-NHCH₂), 4.57 (t, J=5.0Hz, 1H ,-OH), 3.56 (m, J=5.8Hz, 2H ,-CH₂OH),3.47(m,1H,-NHCH(CH₂)₂OH), 3.22 (m, J=2.7,11.1Hz, 1H ,-NHCHH '), 2.84 (m, J=1.6,6.7,11.1Hz, 1H ,-NHCHH '), 1.65 (m, J=6.7Hz, 1H ,-CHH ' CH₂OH), 1.54 (m, J=6.3Hz, 1H ,-CHH ' CH₂OH)；¹³C-NMR(APT)(75MHz, DMSO-d6):δ146.75(C),134.44(C),128.20(C),118.97(CH),110.59(CH),57.97(CH₂), 47.47(CH),43.99(CH₂),36.60(CH₂)。

Synthesize chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7-

To (S) -2- (the chloro- 1,2,3,4- tetrahydropyridines of 6- simultaneously [2,3-b] pyrazine -3- bases) ethanol (5；50g,0.234mol) In CH₂Cl₂Triethylamine (95g, 0.936mol) is added in solution in (500mL).Mixture is stirred until it in environment temperature Uniformly, it is subsequently cooled to 0 DEG C.POCl is added dropwise into reactant mixture₃(54g, 0.351mol), while maintaining temperature at 0-5 DEG C Between.Cooling bath is removed, reactant mixture is then stirred into 2h in environment temperature, disappearing for alcohol is originated while being monitored via HPLC Lose.After the completion of reaction, 1.2M NaHCO are added₃(aq.)(200mL).Separate each layer, and by water layer CH₂Cl₂Extraction.Merge CH₂Cl₂Layer 1M HCl (aq.) (4 × 300mL) extractions, the HCl layers solid NaHCO of merging₃Adjust to pH 8.Gained is mixed Compound CH₂Cl₂Extract (4 × 300mL), then by part CH₂Cl₂Layer dries (Na₂SO₄), filtering, and handled with charcoal (50g).Mixture is stirred into 3h, filtering, then by charcoal CH in environment temperature₂Cl₂(200mL) is washed.The filtrate of merging and wash Wash solution to be concentrated to dryness, and solid residue is dried in vacuo, obtain (4S) -7- chloro- 2,3,4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-endo-methylene group Pyrido [2,3-b] [1,4] diazaIt is pale white crystals solid (30g, 66%).LRMS(m/z):196.1[M+H] +；HRMS(m/z):[M+H]+C₉H₁₁N₃Cl calculated value, 196.0642；Measured value, 196.0637；¹H-NMR(300MHz, DMSO-d6):δ 7.47 (br d, J=4.5Hz, 1H ,-NH), 7.09 (d, J=7.7Hz, 1H), 6.39 (d, J=7.7Hz, 1H), 3.89 (m, J=5.0Hz, 1H, CHNH), 2.95-3.13 (m, 2H ,-NCH₂CH₂CHNH),2.77(m,2H,-NCHH’CHNH), 1.98 (m, J=5.0Hz, 1H ,-NHCHCHH ' CH₂N), 1.86 (m, J=6.9Hz, 1H ,-NHCHCHH ' CH₂N)；¹³C-NMR (APT)(75MHz,DMSO-d6):δ153.45(C),144.50(C),134.32(CH),133.19(C),109.73(CH), 59.88(CH₂),53.07(CH₂),50.08(CH),38.38(CH₂)。

The bicyclic pyridine core of the substitution of synthesis 3

Synthesize fluoro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of the chloro- 8- of (4S) -7-

The fluoro- N'- chloromethyls triethylenediamines of N- double (tetrafluoroborate) (15.20g, 42.9mmol) are added in room temperature Chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7-(7g, 35.8mmol) exists In solution in trifluoromethanesulfonic acid (70mL), 70 DEG C are then heated to, 36h is kept.Then by reactant mixture NaHCO₃Water Solution is neutralized and is extracted with ethyl acetate (3x500mL).The organic layer of merging is washed with water (2x500mL) and salt solution (100mL), Through anhydrous Na₂SO₄Dry, then removal of solvent under reduced pressure, obtain crude residue.Crude mixture is through flash column chromatography (silicon Glue：100-200 mesh, eluant, eluent：1%MeOH/DCM), the chloro- 8- of (4S) -7- fluoro- 2,3,4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-endo-methylene group are obtained Pyrido [2,3-b] [1,4] diaza(200mg, 0.805mmol, yield：2.25%), it is pale solid.(TLC: Eluant, eluent：5% methanol/DCM, R_f:0.4), LCMS (m/z) 214.1 [M+H]⁺。

Iodo- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of the chloro- 8- of (4S) -7-

To chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7-(10g, 51.1mmol) NIS (14.95g, 66.4mmol) is added in the solution of stirring in chloroform (170mL).Reactant mixture is existed 70 DEG C of stirring 1h.Reactant mixture is reached room temperature, (50ml) is diluted with water and CHCl is used₃Extract (2x100ml).What is merged has Machine layer aqueous salt solu-tion and through anhydrous Na₂SO₄Dry, be evaporated under reduced pressure, obtain crude product.Crude product is pure through flash column chromatography Change (silica gel:100-200 mesh), obtain the chloro- 8- of (4S) -7- iodo- 2,3,4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-endo-methylene group pyrido [2,3-b] [1,4] diaza(7.4g, 20.53mmol, 40.2% yield), it is yellow solid (TLC systems：Net EtOAc, (R_f: 0.4).LCMS(m/z)321.96,[M+H]⁺。

Synthesis (4S) -7- chloro- 8- (trifluoromethyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza

Exist in room temperature to cuprous iodide (I) (1.355g, 13.68mmol), potassium tert-butoxide (1.535g, 13.68mmol) Trifluoromethyl trimethylsilane (1.983mL, 13.68mmol), 1,10- phenanthrene is added in the solution of stirring in DMPU (34mL) to cough up Quinoline (2.466g, 13.68mmol) and the iodo- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyridos [2,3-b] of the chloro- 8- of (4S) -7- [1,4] diaza(2.2g,6.84mmol).24h is stirred at room temperature in reactant mixture, then reactant mixture is toppled over Into cold water (210mL), and (2x200mL) is extracted with ethyl acetate.The organic layer of merging is through anhydrous Na₂SO₄Dry, vacuum is dense Contracting, obtains crude compound.Crude product is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：3%CH₂Cl₂/ EtOAc), the chloro- 8- of (4S) -7- (trifluoromethyl) -2,3,4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-endo-methylene group pyrido [2,3-b] [Isosorbide-5-Nitrae] are obtained Diaza(650mg, 2.465mmol, yield：36.0%), it is pale solid (TLC systems：Net EtOAc, R_f:0.5) .LCMS(m/z):264.14[M+H]⁺, Rt=1.78min.

Synthesize (R) -2- ((the chloro- 3- nitropyridines -2- bases of 6-) amino) dimethyl succinate

Under a nitrogen to (R) -2- aminosuccinic acid diformazan ester hydrochlorides (25g, 127mmol) at tetrahydrofuran (THF) In suspension in (130mL) add sodium acid carbonate (21.25g, 253mmol) and 2,6- dichloro-3-nitropyridines (12.21g, 63.3mmol).Reactant mixture is stirred into 16hr at 40 DEG C.Reactant mixture is filtered and washed (3 × 25mL) with EtOAc, Filtrate is concentrated, crude product is obtained, is then added in silicagel column, with (9:1) Hex/EtOAc is eluted.The fraction of collection is steamed Hair, obtains desired product (16g, 49.4mmol, 39.0%), LCMS (m/z) 318.1 [M+H]⁺。

Synthesize (R) -2- (6- chloro-2-oxo -1,2,3,4- tetrahydropyridines simultaneously [2,3-b] pyrazine -3- bases) acetoxymethyl ester

To (R) -2- ((the chloro- 3- nitropyridines -2- bases of 6-) amino) dimethyl succinate (16g, 50.4mmol) in isopropyl Iron (14.06g, 252mmol) is added in solution in alcohol (200mL) and water (60mL) and 40 DEG C are heated to.To reactant mixture Middle addition acetic acid (1.442mL, 25.2mmol), is then heated to 70 DEG C, keeps 1hr.Reactant mixture is cooled to room temperature, passed through Diatomite is filtered and washed (3 × 20mL) with EtOAc, concentrates filtrate and drying.Reacting coarse product must expire from ethyl alcohol recrystallization The product (11.5g, 43.4mmol, 86%) of prestige, LCMS (m/z) 256.1 [M+H]⁺。

Synthesize (R) -2- (the chloro- 1,2,3,4- tetrahydropyridines of 6- simultaneously [2,3-b] pyrazine -3- bases) ethanol

It is mixed in tetrahydrofuran (THF) (12mL) to lithium aluminium hydride reduction (8.54g, 225mmol) in a nitrogen atmosphere at 0 DEG C (R) -2- (6- chloro-2-oxo -1,2,3,4- tetrahydropyridines simultaneously [2,3-b] pyrazine -3- bases) acetoxymethyl ester is added dropwise in suspension The solution of (11.5g, 45.0mmol) in tetrahydrofuran (THF) (60mL).Reactant mixture is heated to 70 DEG C, holding 16hr.Reactant mixture is cooled to 0 DEG C, is quenched with water (8mL), keeps internal temperature to be less than 5 DEG C., will be anti-after addition terminates Mixture is answered to stir 15min.Then, 10mL 15% (W/W) NaOH (aq.) is added, keeps internal temperature to be less than 5 DEG C, knot is added Shu Hou, 15min is stirred by reactant mixture.In order to complete post processing, 12mL water is added, then mixture is stirred at room temperature 1h.Solid is filtered and washed with THF (3x20mL), filtrate and washings are concentrated in vacuo.Crude product is added into silicagel column to be used in combination (3:7) Hex/EtOAc is eluted.The fraction of collection is evaporated, desired product (6g, 27.0mmol, 60.1%), LCMS is obtained (m/z)214.1[M+H]⁺。

Synthesize chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4R) -7-

Under a nitrogen to (R) -2- (the chloro- 1,2,3,4- tetrahydropyridines of 6- simultaneously [2,3-b] pyrazine -3- bases) ethanol (7g, Triethylamine (18.27mL, 131mmol) 32.8mmol) is added in the solution in dichloromethane (DCM) (70mL).By mixture It is stirred at room temperature until it is uniform, is then cooled to 0 DEG C.Then POCl is added dropwise₃(4.58mL, 49.1mmol), maintains temperature Spend for 0 DEG C to 5 DEG C.Reactant mixture is stirred into 2hr at 25 DEG C.After the completion of reaction, 100mL 1.2M NaHCO are added₃(aq.)。 Separate each layer and extract water layer (2x200mL) with DCM.The organic layer of merging is concentrated to dryness.Solid residue is dried in vacuo. Crude product 8g adds to silicagel column and eluted with 70%EtOAc/ petroleum ethers.The fraction of collection is evaporated, desired product is obtained (5.2g, 26.5mmol, 81% yield), LCMS (m/z) 195.9 [M+H]⁺。

The bicyclic pyridine core of the substitution of synthesis 4

Synthesize the chloro- 4- Methyl-3-nitropyridines of 2,6- bis-

Nitric acid (1.5mL, 33.6mmol) is added to sulfuric acid (2.5mL, 46.9mmol) solution of stirring at 0 DEG C under a nitrogen In.Then the chloro- 4- picolines (0.500g, 3.09mmol) of 2,6- bis- are added at 0 DEG C.Then by reactant mixture at 100 DEG C Stir 16hr.Monitored and reacted by TLC.After the completion of, reactant mixture is quenched with trash ice, NH is used₄OH solution is neutralized and filtered Solid, is then dried in vacuo, and obtains desired product (0.300g, 1.443mmol, 46.8% yield), it is faint yellow solid, LCMS(m/z)206.8[M+H]⁺。

Synthesize (S) -2- ((the chloro- 4- Methyl-3-nitropyridines -2- bases of 6-) amino) dimethyl succinate

At 0 DEG C under a nitrogen by the chloro- 4- Methyl-3-nitropyridines (300mg, 1.449mmol) of 2,6- bis- and sodium acid carbonate (243mg, 2.90mmol) is at tetrahydrofuran (THF) (20mL)) in suspension add to (S) -2- aminosuccinic acid dimethyl ester salt In hydrochlorate (430mg, 2.174mmol).Then reactant mixture is stirred into 24hr at 65 DEG C.Monitored and reacted by TLC.Will reaction Material is filtered and washed (2 × 30mL) with EtOAc.Be concentrated under reduced pressure filtrate, obtains thick material.Crude product is added into neutral alumina In post and with Hex/EtOAc (9:1) elute.The fraction of collection is concentrated under reduced pressure, obtain desired product (250mg, 0.742mmol, 51.2% yield), it is yellow colloidal liquid, LCMS (m/z) 339.1 (M+H)⁺。

Synthesize (S) -2- (the chloro- 8- methyl -2- oxos -1,2,3,4- tetrahydropyridines of 6- simultaneously [2,3-b] pyrazine -3- bases) acetic acid Methyl esters

To (S) -2- ((the chloro- 4- Methyl-3-nitropyridines -2- bases of 6-) amino) dimethyl succinate in 40 DEG C of stirrings (6.0g, 18.09mmol) and iron (5.05g, 90mmol) add second in the suspension in isopropanol (80mL) and water (20mL) Sour (1.553mL, 27.1mmol).Reactant mixture is stirred into 1hr at 80 DEG C.Monitored and reacted by TLC.Reactant mixture is cold But to room temperature, then it is quenched and is extracted with EtOAc with saturated sodium bicarbonate solution.Organic layer aqueous salt solu-tion simultaneously uses sulfuric acid Sodium drying and dehydrating, filters and evaporates, and obtains desired product (4.0g, 14.32mmol, 79% yield), LCMS (m/z) 269.9 [M+H]⁺。

Synthesize (S) -2- (the chloro- 8- methyl isophthalic acids of 6-, 2,3,4- tetrahydropyridines simultaneously [2,3-b] pyrazine -3- bases) ethanol

To control the speed that gas is escaped, to the aluminium chloride (0.173g, 1.298mmol) stirred under a nitrogen in tetrahydrochysene furan It is added dropwise in the solution muttered in (THF) (2.5mL) in solution of the 2M lithium aluminium hydride reductions (2.220mL, 4.44mmol) in THF.This is obtained To aluminum hydride (AlH3) THF solution.In a single flask, (S) -2- (chloro- 8- methyl -2- of 6- are prepared under a nitrogen Oxo -1,2,3,4- tetrahydropyridines simultaneously [2,3-b] pyrazine -3- bases) methyl acetate (0.250g, 0.927mmol) is in tetrahydrofuran (THF) solution in (5mL), is added dropwise hydrogenation aluminum solutions at -78 DEG C, lasts 15 minutes thereto.After addition terminates, cooling is removed Bath, and cause reactant mixture to be warmed to environment temperature.Monitored and reacted by TLC.At 0 DEG C by reactant mixture 10%NaOH Solution is quenched, and then stirs 1hr and is extracted with EtOAc.EtOAc layers of priority are dried with water and aqueous salt solu-tion and with sodium sulphate Dehydration, filters and concentrates, obtain desired product (150mg, 0.407mmol, 43.9% yield), it is faint yellow solid, LCMS(m/z)228.2[M+H]⁺。

Synthesize chloro- 9- methyl -2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazines of (4S) -7- It is miscellaneous

To (S) -2- (the chloro- 8- methyl isophthalic acids of 6-, 2,3,4- tetrahydropyridines simultaneously [2,3-b] pyrazine -3- bases) ethanol (1.8g, HBr (4mL, 35.4mmol) is added in 7.91mmol), reactant mixture is stirred into 18hr at 90 DEG C.Monitored and reacted by TLC. After the completion of, reactant mixture is quenched with saturated sodium bicarbonate solution and extracted with EtOAc.EtOAc layers of priority water and salt solution Solution washs and uses sodium sulphate drying and dehydrating, filters and concentrates, obtains crude product.Crude product is added in neutral alumina and is used in combination 20%EtOAc/ Hex.The fraction of collection is evaporated, obtaining desired product, (0.900g, 4.27mmol, 54.0% are received Rate), it is faint yellow solid, LCMS (m/z) 210.2 [M+H]⁺。

Chlorination coupling reaction

Synthesize (4S) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 7- formic acid first Ester

To chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7-(5g, TEA (17.77mL, 127.77mmol) and Pd 25.55mmol) is added in the solution of the degassing in anhydrous MeOH (250ml) (dppf)Cl₂(934mg, 1.2755mmol) and reactant mixture is stirred under 300psi nitric oxide atmosphere at 110 DEG C 20h.Suspension is cooled to room temperature and mixture is concentrated under reduced pressure, crude compound is obtained.Crude mixture is through quick post color Spectrum purifying (100-200 silica gel is eluted with 2% methanol/DCM), obtains (4S) -2,3,4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-endo-methylene group pyridine And [2,3-b] [1,4] diaza- 7- methyl formates (3g, 13.68mmol, 53.5% yield), it is Light brown solid (TLC System:5% methanol/DCM, R_fValue：0.2), LCMS (m/z) 220.3 [M+H]⁺。

Synthesize (4S) -7- (6- picoline -3- bases) -8- (trifluoromethyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza

To (4S) -7- chloro- 8- (trifluoromethyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1,4] Diaza(350mg, 1.328mmol), (6- picoline -3- bases) boric acid (273mg, 1.991mmol) and K₃PO₄ Pd is added in the solution of the degassing of (1096mg, 3.98mmol) in 1,4- dioxanes (10mL) and water (3mL)₂(dba)₃ (122mg, 0.133mmol) and x-phos (633mg, 1.328mmol).Reactant mixture is stirred into 7h at 100 DEG C.So that reaction Mixture reaches room temperature and is diluted with water (70mL), is extracted with ethyl acetate (3 × 150mL).The organic layer of merging is through anhydrous Na₂SO₄Dry and be concentrated under reduced pressure, obtain crude product.Crude mixture grinds (3x25mL) and vacuum drying with petroleum ether, obtains (4S) -7- (6- picoline -3- bases) -8- (trifluoromethyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza(370mg, 1.040mmol, yield：78%), it is pale solid (TLC systems:5% methanol/ EtOAc,Rf:0.3).LCMS(m/z)320.90[M+H]⁺。

Synthesis (4S) -8- fluoro- 7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza

By K₃PO₄(397mg, 1.872mmol) adds to the fluoro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene group pyrroles of the chloro- 8- of (4S) -7- Pyridine simultaneously [2,3-b] [1,4] diaza(200mg, 0.936mmol) ＆ (6- picoline -3- bases) boric acid (167mg, 1.217mmol) in the solution of the stirring in the dioxane of Isosorbide-5-Nitrae-(10mL) and water (1mL), then deaerate 15min, then successively Add x-phos (44.6mg, 0.094mmol) and Pd₂(dba)₃(42.9mg,0.047mmol).Reactant mixture is added at 90 DEG C Hot 2h 45min.Reactant mixture is cooled to room temperature, filtered by Celite pad, (10mL x2) is washed with ethyl acetate, so Removal of solvent under reduced pressure afterwards.(20mL x2) is extracted with ethyl acetate in organic compound, is washed with water (2 × 20mL) and salt solution (20mL) Wash.Extract is through anhydrous Na₂SO₄Dry, solvent is removed in vacuum, crude compound is obtained.Crude compound is purified through column chromatography, Eluted using 100-200 silica gel and with 1%MeOH/DCM, obtain the fluoro- 7- of (4S) -8- (6- picoline -3- bases) -2,3,4,5- Tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(220mg, 0.705mmol, 75% yield), it is ash White solid, LCMS (m/z) 271.15 [M+H]⁺。

Synthesize (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza

To chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7-(30g, 153mmol), (2- picoline -4- bases) boric acid (25.2g, 184mmol) and tripotassium phosphate (65.1g, 307mmol) are in 1- fourths In alcohol (300mL) and the solution of the degassing in water (50.0mL) add three (dibenzalacetone) two palladiums (0) (7.02g, 7.67mmol) with dicyclohexyl (2', 4', 6'- triisopropyl-[1,1'- biphenyl] -2- bases) phosphine (7.31g, 15.33mmol).Will Reactant mixture heats 3h at 100 DEG C.N-butanol solvent is evaporated under reduced pressure.Gained residue diluted with water (200mL) is simultaneously extracted with DCM Take (2x400ml).The organic layer water of merging, salt water washing, it is dried over sodium sulfate and solvent is evaporated under reduced pressure, obtain crude product. Crude product ether and pentane (1:1) grind 3 times (3X250mL, obtains (4S) -7- (2- picoline -4- bases) -2,3,4, 5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(25g, 99.2mmol, 62%), it is canescence Solid (TLC:10%MeOH/EtOAc R_f:0.2), LCMS (m/z) 252.9 [M+H]⁺。

¹H NMR(400MHz,DMSO-d6):δ 8.45 (dd, J=5.2,0.8Hz, 1H), 7.73 (dt, J=1.4, 0.7Hz, 1H), 7.67-7.60 (m, 1H), 7.24-7.16 (m, 2H), 7.10 (d, J=7.7Hz, 1H), 3.93 (td, J=5.0, 2.5Hz,1H),3.19-2.98(m,2H),2.92-2.71(m,2H),2.50(s,3H),2.11-1.96(m,1H),1.94- 1.81(m,1H)。

Synthesize (4S) -7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza

To chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7-(10g, 51.1mmol), (6- picoline -3- bases) boric acid (10.50g, 77mmol) and tripotassium phosphate (21.70g, 102mmol) be 1, In 4- dioxanes (100mL) and the solution of the degassing in water (20.0mL) add three (dibenzalacetone) two palladiums (0) (4.68g, 5.11mmol) with x-phos (4.87g, 10.22mmol).Reactant mixture is heated into 8h at 100 DEG C.Solvent is evaporated under reduced pressure；Will The residue diluted with water (200mL) of acquisition is simultaneously extracted (2x100ml) with DCM.The organic layer water of merging, salt water washing, warp Sodium sulphate is dried and solvent is evaporated under reduced pressure, and obtains crude product.Crude product ether and pentane (1:1) grind 3 times (3X100mL), obtains (4S) -7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-endo-methylene group pyrido [2,3-b] [1,4] diaza(11.2g, 44.4mmol, 65%), it is pale solid (TLC:10%MeOH/EtOAc R_f: 0.3).LCMS(m/z):253.1 [M+H], Rt=2.86min.

¹H NMR(400MHz,CDCl₃):δ ppm 8.97 (d, J=2.19Hz, 1H), 8.06 (dd, J=8.11,2.41Hz, 1H), 7.28-7.11 (m, 1H), 6.94 (d, J=7.89Hz, 1H), 6.74 (d, J=7.67Hz, 1H), 5.21 (s, 1H), 4.08–3.98(m,1H),3.36-3.12(m,3H),2.99-2.89(m,1H),2.56(s,3H),2.17-2.10(m,2H)。

Synthesize (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza

By chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7-(20g, 102mmol), (3- (trifluoromethyl) phenyl) boric acid (29.1g, 153mmol) and Cs₂CO₃(100g, 307mmol) is in 1,4- bis- Evil Suspension in alkane (100mL) and water (10mL) stirs and uses argon gas to be deaerated 15 minutes in room temperature.Then, by acid chloride (II) (0.574g, 2.56mmol) and 2- dicyclohexyls phosphino- -2 ', 4 ', 6 '-tri isopropyl biphenyl (2.437g, 5.11mmol) is added to In reactant mixture.Then reactant mixture is stirred into 2hr at 110 DEG C.Reactive material is filtered and concentrated through diatomite.By remnants Thing is with EtOAc dilutions and successively uses saturation NaHCO₃And aqueous salt solu-tion, sodium sulphate drying and dehydrating is then used, filters and steams Hair.Crude product adds to silicagel column and with Hex/EtOAc (1:1) elute.The fraction of collection is evaporated, (4S) -7- (3- (three are obtained Methyl fluoride) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(18g, 58.3mmol, 57.0% yield), it is white solid, LCMS (m/z) 306.1 (M+H)⁺。

Synthesize (4S) -9- methyl -7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza

By chloro- 9- methyl -2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7-(0.900g, 4.29mmol), (6- picoline -3- bases) boric acid (0.882g, 6.44mmol) and tripotassium phosphate (2.73g, 12.88mmol) suspension in 1,4- dioxanes (14.4mL) ＆ water (3.6mL) stirs and uses argon gas to be deaerated 15 points in room temperature Clock.Then by Pd₂(dba)₃(0.393g, 0.429mmol) and X-Phos (0.409g, 0.858mmol) add to reactant mixture In.Reactant mixture is stirred into 16hr at 90 DEG C.Pass through TLC monitoring reactions (50%EtOAc/ hexanes).Reactant mixture is cold But to room temperature, then filter and washed with EtOAc through diatomite.Filtrate is concentrated, is then dissolved with EtOAc.EtOAc layers are successively used Water and aqueous salt solu-tion, it is then dried over sodium sulfate, filter and concentrate, obtain thick material (4S) -9- methyl -7- (6- methyl Pyridin-3-yl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(0.700g, 2.60mmol, 60.7% yield), it is pale solid, LCMS (m/z) 267.0 [M+H]⁺。

Synthesize (2R) -2- ethyls -4- ((4S) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine- 7- bases) morpholine

By chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7- (900mg, 4.60mmol), (R) -2- ethyl morpholines (1060mg, 9.20mmol) and KOtBu (1032mg, 9.20mmol) 1, Suspension in 2- dimethoxy-ethanes (DME) (20mL) stirs and in room temperature degassing 15min, adds (1,3- bis- under a nitrogen (2,6- diisopropyl phenyls) -4,5- glyoxalidine -2- subunits) chlorine) (3- phenyl allyls) palladium (2) (120mg, 0.184mmol).Reactant mixture is stirred into 12hr at 90 DEG C.Reactive material filters through Celite pad and concentrates filtrate.Reaction is mixed Compound is diluted with water, and is extracted (2 × 20mL) with EtOAc.Organic layer priority water and aqueous salt solu-tion are simultaneously dry with sodium sulphate Dry dehydration.Organic layer is concentrated under reduced pressure.Crude compound is purified (neutral alumina) through column chromatography.Product with 20% ethyl acetate/ Hex.The fraction of collection is evaporated under reduced pressure and in high vacuum dry, obtains (2R) -2- ethyls -4- ((4S) -2,3,4,5- Tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 7- bases) (750mg, 2.73mmol, 59.4% are received morpholine Rate), it is faint yellow solid, LCMS (m/z) 275.3 [M+H]⁺。

Synthesize (2S, 6S) -2,6- dimethyl -4- ((4S) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 7- bases) morpholine

By chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7- (0.800g, 4.09mmol), (2S, 6S) -2,6- thebaines (0.942g, 8.18mmol) and KOtBu (0.918g, 8.18mmol) stir and deaerated 15 points in room temperature with argon gas in the suspension in 1,2- dimethoxy-ethanes (DME) (30mL) Clock.Then by (1,3- bis- (2,6- diisopropyl phenyls) -4,5- glyoxalidine -2- subunits) chlorine) (3- phenyl allyls) palladium (II) (0.106g, 0.164mmol) is added in reactant mixture.Then reactant mixture is stirred into 16hr at 90 DEG C.Pass through TLC Monitoring reaction.After the completion of, reactive material is filtered through diatomite, and be then concentrated under reduced pressure filtrate.By gained reactant mixture EtOAc Dilute and priority water and aqueous salt solu-tion simultaneously use sodium sulphate drying and dehydrating, filter and evaporate and obtain crude product.By crude product Add in neutral alumina and with (1:2) EtOAc/ Hex.The fraction of collection is evaporated, (2S, 6S) -2,6- diformazans are obtained Base -4- ((4S) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas- 7- bases) morpholine (0.780g, 2.79mmol, 68% yield), it is pale solid, LCMS (m/z) 275.3 [M+H]⁺。

Synthesize (2R, 6R) -2,6- dimethyl -4- ((4S) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 7- bases) morpholine

By chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7- (0.800g, 4.09mmol), (2R, 6R) -2,6- thebaines (0.942g, 8.18mmol) and KOtBu (0.918g, Stirred and with argon gas in room temperature degassing 15 in 1,2- dimethoxy-ethanes (DME) (30mL) in suspension in 8.18mmol) Minute.Then by (1,3- bis- (2,6- diisopropyl phenyls) -4,5- glyoxalidine -2- subunits) chlorine) (3- phenyl allyls) palladium (2) (0.106g, 0.164mmol) is added in reactant mixture.Then reactant mixture is stirred into 16hr at 90 DEG C.Pass through TLC Monitoring reaction (TLC:100%EtOAcR_fValue：0.2).Reactive material filters through diatomite and is completely distilled off out solvent.Reaction is mixed Compound is with EtOAc dilutions and priority water and aqueous salt solu-tion and uses sodium sulphate drying and dehydrating, filters and evaporates and is slightly produced Thing.Crude product is added in neutral alumina and with (1:1) EtOAc/ Hex.The fraction of collection is evaporated, obtain (2R, 6R) -2,6- dimethyl -4- ((4S) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas-7- Base) morpholine (0.800g, 2.75mmol, 67.3% yield), it is pale solid, LCMS (m/z) 275.0 [M+H]⁺。

Synthesize (2S, 6R) -2,6- dimethyl -4- ((4S) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 7- bases) morpholine

By chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7- (1.00g, 5.11mmol), (2S, 6R) -2,6- thebaines (1.177g, 10.22mmol) and KOtBu (1.147g, 10.22mmol) suspension in 1,2- dimethoxy-ethanes (DME) (20mL) stirs and uses argon gas to be deaerated 15 points in room temperature Clock.Then by (1,3- bis- (2,6- diisopropyl phenyls) -4,5- glyoxalidine -2- subunits) chlorine) (3- phenyl allyls) palladium (II) (0.133g, 0.204mmol) is added in reactant mixture.Reactant mixture is stirred into 16hr at 90 DEG C.Reactive material is passed through Diatomite filtering and evaporation solvent.By reactant mixture EtOAc dilutions and priority water and aqueous salt solu-tion, then through sulphur Sour sodium is dried, and is filtered and is evaporated, obtains thick material (2S, 6R) -2,6- dimethyl -4- ((4S) -2,3,4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-bridge Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 7- bases) morpholine (0.820g, 2.86mmol, 56.0% yield).Crude product Add in neutral alumina and use 20%EtOAc/ Hex, obtain (2S, 6R) -2,6- dimethyl -4- ((4S) -2,3,4, 5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 7- bases) morpholine (0.820g, 2.86mmol, 56.0% yield), it is pale solid, LCMS (m/z) 275.2 [M+H]⁺。

Synthesize 2- cyclopropyl -4- ((4S) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazines It is miscellaneous- 7- morpholines

Under agitation at 0 DEG C to the chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyridos [2,3-b] [1,4] of (4S) -7- Diaza2- cyclopropyl morpholines are added in the solution of (1g, 5.11mmol) in 1,2- dimethoxy-ethanes (DME) (15mL) (0.650g, 5.11mmol) and potassium tert-butoxide (0.574g, 5.11mmol).By solution and then the 15min that deaerates, and add cinnamoyl Base chlorine [1,3- bis- (diisopropyl phenyl) -2- imidazolidinyls subunit] Pd (II) (3.32g, 5.11mmol).Then it is reaction is mixed Compound stirs 16h at 90 DEG C.Reactant mixture is gone out with 15ml water quenchings and uses 15ml ethyl acetate to extract.Organic layer is through sodium sulphate Dry and be concentrated under reduced pressure, obtain crude compound.Crude product is added into 100-200 silicagel columns and eluted with 2%DCM/MeOH, is obtained To product 2- cyclopropyl -4- ((4S) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas-7- Base) morpholine (1g, 3.49mmol, 68.3% yield), it is pale solid, LCMS (m/z) 287.2 [M+H]⁺。

Synthesize 2,2- dimethyl -4- ((4S) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine- 7- bases) morpholine

By chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7-(1.0g, 5.11mmol), 2,2- thebaines (1.177g, 10.22mmol) and KOtBu (1.147g, 10.22mmol) are in 1,2- diformazans Stir and deaerated 15 minutes in room temperature with argon gas in suspension in epoxide ethane (DME) (20mL).Then incite somebody to action (1,3- bis- (2, 6- diisopropyl phenyls) -4,5- glyoxalidine -2- subunits) chlorine) (3- phenyl allyls) palladium (2) (0.133g, 0.204mmol) Add to reactant mixture.Reactant mixture is stirred into 16hr at 90 DEG C.Reactive material is through diatomite filtering and evaporation solvent.Will be anti- Answer mixture EtOAc dilutions and priority water and aqueous salt solu-tion.Solution over sodium sulfate is dried, filters and evaporates, obtain To crude product.Crude product is added into neutral alumina column and eluted with DCM, 2,2- dimethyl -4- ((4S) -2,3,4,5- are obtained Tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 7- bases) morpholine (0.800g, 2.67mmol, 52.3% Yield), it is white solid, LCMS (m/z) 275.0 [M+H]⁺。

Synthesis (4S) -7- (3- (trifluoromethyl) piperidin-1-yl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza

At 30 DEG C to chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7-(500mg, 2.56mmol), 3- (trifluoromethyl) piperidines (783mg, 5.11mmol (S- isomers, the mapping containing inequality Body mixture)) add in the solution of degassing in 1,2- dimethoxy-ethanes (15mL) potassium tert-butoxide (574mg, 5.11mmol) with Umicore catalyst (33.2mg, 0.051mmol).Reactant mixture is heated into 17h at 80 DEG C.Reaction is mixed Compound is cooled to room temperature and poured into cold water (70mL), is extracted with ethyl acetate (2x150mL).The organic layer of merging is through anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain crude product.Crude mixture is through flash column chromatography (silica gel：100-200 Mesh, eluant, eluent：1 to 3% methanol/ethyl acetate), 400mg (4S) -7- (3- (trifluoromethyl) piperidin-1-yl) -2,3,4 is obtained, 5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaChiral HPLC indicates 53:44 mixture of enantiomers, The chiral preparation HPLC purifying of mixture of enantiomers of the inequality, obtains peak (the chiral preparative condition of two separation： The 0.5%DEA/ methanol of 4g-40%-100 bars 10, chiralpak, AD-H (4.6mm*250mm)), it is the peak most eluted soon: 210mg (peak-I, 0.673mmol, 15% yield), it is white solid (TLC:10%MeOH/EtOAc R_f:0.4) it is, and most slow The peak eluted:310mg (peak-II, 310mg, 0.993mmol, 22%), it is white solid (TLC:10%MeOH/EtOAc, R_f:0.4), LCMS (m/z) 313.2 [M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ ppm 6.92 (d, J=8.33Hz, 1H), 6.56 (d, J=4.38Hz, 1H), 5.83 (d, J=8.11Hz, 1H), 4.43 (d, J=12.28Hz, 1H), 3.93 (d, J=12.28Hz, 1H), 3.87-3.72 (m, 1H),3.10–2.83(m,2H),2.82-2.71(m,1H),2.70-2.55(m,3H),2.46-2.18(m,1H),2.11–1.86 (m,2H),1.86-1.65(m,2H),1.55-1.32(m,2H)。

At 30 DEG C to chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7-(500mg, 2.56mmol), (R- isomers, the mixture of enantiomers containing inequality) 3- (trifluoromethyl) piperidines In the solution of the degassing of (783mg, 5.11mmol) in 1,2- dimethoxy-ethanes (15mL) add potassium tert-butoxide (574mg, 5.11mmol) with Umicore catalyst (33.2mg, 0.051mmol).Reactant mixture is heated into 17h at 80 DEG C.Reaction is mixed Compound is cooled to room temperature and poured into cold water (70mL), is extracted with ethyl acetate (2x150mL).The organic layer of merging is through anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain crude product.Crude mixture is through flash column chromatography (silica gel：100-200 Mesh, eluant, eluent：1 to 3% methanol/ethyl acetate), obtain (4S) -7- (3- (trifluoromethyl) piperidin-1-yl) -2,3,4,5- tetra- Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza500mg compounds are separated after column chromatography, chiral HPLC refers to Show 36:63 mixture of enantiomers, the chiral preparation HPLC purifying of mixture of enantiomers of the inequality, two separation of acquisition Peak (chiral preparative condition：4g-40%-100 bar 100.5%DEA/ methanol, chiralpak, AD-H (4.6*250) mm5u), For the peak most eluted soon:140mg (peak-I), and the peak most eluted slowly:310mg (peak-II, 310mg, 0.993mmol, 22%), it is white solid (TLC:10%MeOH/EtOAc, R_f:0.4), LCMS (m/z) 313.2 [M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ ppm 6.92 (d, J=8.33Hz, 1H), 6.56 (d, J=4.60Hz, 1H), 5.83 (d, J=8.33Hz, 1H), 4.41 (dt, J=12.50,1.86Hz, 1H), 3.93 (d, J=12.50Hz, 1H), 3.81 (td, J=4.88,2.74Hz, 1H), 3.07-2.88 (m, 2H), 2.80-2.55 (m, 4H), 2.48-2.27 (m, 1H), 2.01- 1.88(m,2H),1.85-1.65(m,2H),1.52-1.38(m,1H),1.36-1.20(m,1H)。

Modify the pyridine aryl-linking compound of 3- substitutions

Synthesis (4S) -8- bromo- 7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza

It is sub- to (4S) -7- (6- picoline -3- the bases) -2,3,4,5- tetrahydrochysene -1,4- bridges stirred under a nitrogen at 20 DEG C Picoline simultaneously [2,3-b] [1,4] diazaNBS is added in the solution of (1g, 3.96mmol) in chloroform (10mL) (0.776g,4.36mmol).Reactant mixture is stirred into 2h at 70 DEG C.So that reactant mixture reaches room temperature and uses saturation NaHCO₃It is quenched, uses CHCl₃Extract (2x100ml).The organic layer aqueous salt solu-tion of merging and through anhydrous Na₂SO₄Dry, It is evaporated under reduced pressure, obtains crude compound.Crude mixture is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：3% MeOH/EtOAc), the bromo- 7- of (4S) -8- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-endo-methylene group pyrido are obtained [2,3-b] [1,4] diaza(650mg, 1.962mmol, yield：49.5%), it is yellow solid (TLC systems:10% MeOH/EtOAc, R_f:0.4), LCMS (m/z) 331.1 [M+H]⁺。

Synthesis (4S) -8- bromo- 7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza

It is sub- to (4S) -7- (2- picoline -4- the bases) -2,3,4,5- tetrahydrochysene -1,4- bridges stirred under a nitrogen at 25 DEG C Picoline simultaneously [2,3-b] [1,4] diazaNBS is added in the solution of (10g, 39.6mmol) in chloroform (150mL) (7.76g,43.6mmol).Reactant mixture is stirred into 2h at 70 DEG C.So that reactant mixture reaches room temperature, and reaction is mixed Thing saturation NaHCO₃(100ml) is quenched and uses CHCl₃Extract (2x150ml).The organic layer aqueous salt solu-tion of merging is simultaneously passed through Na₂SO₄Dry.Then it is evaporated under reduced pressure, obtains crude compound.Crude product is obtained through purification by flash chromatography (100-200 mesh) The bromo- 7- of (4S) -8- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous(4.5g, 13.27mmol, 33.5% yield), it is yellow solid (TLC systems:10%MeOH/EtOAC, R_f0.3), LCMS(m/z)330.9[M+H]⁺。

Synthesize (4S) -8- methyl -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza

To the bromo- 7- of (4S) -8- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza(1g, 3.02mmol), methyl-boric acid (0.271g, 4.53mmol) and K₃PO₄(2.493g, Solid Pd 9.06mmol) is added in the solution of the degassing in 1,4- dioxanes (20mL) and water (5mL)₂(dba)₃(0.276g, 0.302mmol) with X-Phos (0.288g, 0.604mmol).Reactant mixture is stirred into 15hr at 100 DEG C.By TLC and LCMS monitoring reaction (TLC systems:- 5% methanol/ethyl acetate, R_f:0.3).(150mL) is diluted with water simultaneously in reactant mixture It is extracted with ethyl acetate (3 × 350mL).Organic layer is through anhydrous Na₂SO₄Dry, be concentrated under reduced pressure, obtain half pure compound.Will be thick Product adds to silicagel column and eluted with DCM/EtOAc.The fraction of collection is concentrated, (4S) -8- methyl -7- (2- methyl pyrroles are obtained Pyridine -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(0.45g, 1.436mmol, 47.6% yield), LCMS (m/z) 267.0 [M+H]⁺。

Synthesize (4S) -8- methyl -7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza

To the bromo- 7- of (4S) -8- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza(1g, 3.02mmol), methyl-boric acid (0.271g, 4.53mmol) and K₃PO₄(2.493g, 9.06mmol) Pd is added in the solution of the degassing in 1,4- dioxanes (20mL) and water (5mL)₂(dba)₃(0.276g, 0.302mmol) with x-phos (0.288g, 0.604mmol).Reactant mixture is stirred into 15h at 100 DEG C.By reactant mixture Room temperature is cooled to, (150mL) is diluted with water and is extracted with ethyl acetate (3 × 350mL).The organic layer of merging is through anhydrous Na₂SO₄ Dry and be concentrated under reduced pressure, obtain crude compound.Crude mixture is through flash column chromatography (silica gel：100-200 mesh, with 3% CH₂Cl₂/ EtOAc is eluted), obtain (4S) -8- methyl -7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-bridge methylene Yl pyridines simultaneously [2,3-b] [1,4] diaza(550mg, 2.044mmol, 67.7%), it is yellow solid (TLC systems: 5% methanol/ethyl acetate, R_f:0.3), LCMS (m/z) .267.3 [M+H]⁺。

Synthesize (4S) -7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 8- formonitrile HCNs

In room temperature to the bromo- 7- of (4S) -8- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza(600mg,1.812mmol)、Zn(CN)₂(638mg, 5.43mmol) and Zn (OAc)₂ Pd is added in the solution of (199mg, 1.087mmol) in N,N-dimethylformamide (DMF) (12mL)₂(dba)₃(332mg, 0.362mmol) with DPPF (402mg, 0.725mmol).Reactant mixture is stirred into 3h at 100 DEG C, it is reached room temperature, (30mL) is diluted with water, is extracted with ethyl acetate (3 × 40mL).The organic layer of merging is through anhydrous Na₂SO₄Dry and be concentrated under reduced pressure, Obtain crude compound.Crude compound is through flash column chromatography (silica gel：100-200 mesh), obtain (4S) -7- (6- methyl Pyridin-3-yl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 8- formonitrile HCNs (350mg, 1.262mmol, yield：69.7%), it is yellow solid (TLC systems:10%MeOH/EtOAc, R_f:0.3), LCMS (m/z) 278.2,[M+H]⁺。

Synthesis (4S) -8- chloro- 7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza

In room temperature to (4S) -7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaN-chloro-succinimide is added in the solution of the stirring of (1g, 3.96mmol) in chloroform (15mL) (NCS,1.058g,7.93mmol).Reactant mixture is stirred into 3h at 70 DEG C, it is reached room temperature, is diluted with water (20ml) and use CH₂Cl₂Extract (2x20ml).The organic layer of merging salt water washing, it is then dried over sodium sulfate, it is evaporated under reduced pressure, Obtain crude product.Crude product is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：3%MeOH/EtOAc), obtain The chloro- 7- of (4S) -8- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous(300mg, 1.032mmol, 26.0% yield), it is pale solid (TLC systems:10%MeOH/EtOAc, R_f0.4), LCMS (m/z) 287.11 [M+H]⁺。

Synthesis (4S) -8- chloro- 7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza

In room temperature to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza(2.5g, 9.91mmol) is in CHCl₃1- chlorine pyrrolidines -2 are added in the solution of stirring in (25mL), 5- diketone (1.59g, 11.9mmol).Reaction solution is set to stir 2h at 60 DEG C.Reaction solution is in CHCl₃And H₂Distributed between O.Point From organic layer, through Na₂SO₄Dry, be concentrated in vacuo and be applied directly to silicagel column, use EtOAc/EtOH (3:1) as elution Agent, obtains (1.5g, 52%), it is yellow colored foam.LCMS(m/z)287[M+H]⁺。

Synthesize (4S) -7- chloro- N- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine- 5 (2H)-formamides

Under a nitrogen at 0 DEG C to the chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyridos [2,3-b] [1,4] of (4S) -7- DiazaIn the agitating solution of (1.2g, 6.13mmol) in tetrahydrofuran (THF) (50mL) add 3- (pyridine -2- bases) - 2H- pyridos [1,2-a] [1,3,5] triazine -2,4 (3H)-diketone (1.768g, 7.36mmol).By reactant mixture at 80 DEG C Stir 16hr, be subsequently poured into cold water (100mL) and be extracted with ethyl acetate (200mL), organic layer in succession with water (70mL) and Salt solution (70mL) is washed, under reduced pressure through anhydrous sodium sulfate drying.Crude residue is purified through column chromatography, uses silica gel (100- 200 mesh) and 30% to 70% gradient mixture as eluant, eluent, obtain 1.3g (66%) title compound, its be canescence thing Matter, LCMS (m/z) 316.2 (M+H)⁺。

Synthesize (4R) -7- chloro- N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine- 5 (2H)-formamides

Stirred under a nitrogen at 0 DEG C to add in pyrazine -2- formic acid (800mg, 6.45mmol) DPPA (3548mg, 12.89mmol) with solution of the triethylamine (4.49mL, 32.2mmol) in tetrahydrofuran (THF) (30mL).By reactant mixture Stir and warm to room temperature, keep 2h.Then, addition (4R) -7- chloro- 2,3,4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-endo-methylene group pyrido [2, 3-b] [1,4] diaza(1009mg,5.16mmol).Reactant mixture is stirred into 16hr at 90 DEG C.Reactant mixture is used Water dilutes (100mL) and is extracted with ethyl acetate (200mL X2).Organic layer is through anhydrous Na₂SO₄Dry, be concentrated under reduced pressure, obtain half Pure compound.Crude product adds to silicagel column and eluted with Hex/EtOAc.Collect fraction obtain desired product (1.4g, 3.71mmol, 58%), LCMS (m/z) 317.2 (M+H)⁺。

Synthesize (4S) -7- chloro- N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine- 5 (2H)-formamides

DIPEA (31.9mL, 183mmol) and DPPA (15.09g, 54.8mmol) are successively added into nicotinic acid in room temperature In the solution of the stirring of (4.5g, 36.6mmol) in tetrahydrofuran (THF) (60mL) and stir 2h.Then it is (4S) -7- is chloro- 2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(5.01g, 25.6mmol) adds to reaction Mixture, is then stirred to 80 DEG C, keeps 16h.Reactant mixture is cooled to room temperature, (60mL) is diluted with water, acetic acid second is used Ester extracts (2X50mL), with salt water washing (50mL).Organic layer is separated, it is dried over sodium sulfate, filter and concentrate, obtain roughization Compound.Crude compound is purified through column chromatography, and using silica gel (100-200 mesh), 1% methanol/DCM obtains desired product (4g, 12.03mmol, 32.9% yield), LCMS:(m/z)316.2(M+H)⁺。

Synthesize (4S) -7- chloro- N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine- 5 (2H)-formamides

In the molten of 0 DEG C of stirring to pyrazine -2- formic acid (4.08g, 32.9mmol) in tetrahydrofuran (THF) (100ml) Diphenyl phosphate azide (14.19ml, 65.8mmol) and TEA (22.94ml, 165mmol) are successively added in liquid.Solution is warmed To room temperature and continue stir 3h.Then, addition solid (4S) -7- chloro- 2,3,4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-endo-methylene group pyrido [2, 3-b] [1,4] diaza(5.6g, 28.6mmol) and heat the mixture to backflow.By reactant mixture stirring 2h backflows Under, it is subsequently cooled to room temperature and continues to be stirred overnight.Next day, mixture is diluted with water and extracted with EtOAc (three times).Merge EtOAc extracts through Na₂SO₄It is dried and concentrated, obtains crude product.Dark residue is through silica gel chromatography:330g posts, 100ml/min, 0-25%EtOAc/MeOH, last 30min.Merge the fraction containing product, obtain desired product, it is yellow Grease.Et is added into the grease₂O (50mL) is simultaneously concentrated under reduced pressure.This causes product to crystallize, and obtains faint yellow solid (6.9g, 76% yield).¹H NMR(400MHz,CDCl₃) δ=12.61 (s, 1H), 9.47 (d, J=1.4Hz, 1H), 8.68- 7.99 (m, 2H), 7.50 (d, J=8.0Hz, 1H), 6.96 (d, J=8.0Hz, 1H), 5.65 (dd, J=6.0,3.2Hz, 1H), 3.30-3.15 (m, 1H), 3.11 (dt, J=12.1,2.1Hz, 1H), 3.00 (dd, J=12.1,3.2Hz, 1H), 2.39-2.22 (m,1H),2.11–1.97(m,2H)；LCMS(m/z)316.9(M+H)⁺。

Synthesize the chloro- 8- of (4S) -7- fluoro- N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides

Triethylamine (1.63mL, 11.737mmol) and triphosgene (696mg, 2.347mmol) are successively added into (4S) -7- Chloro- fluoro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of 8-(500mg,2.347mmol) In solution in THF (20mL), 30min is then stirred at room temperature and 3- aminopyridines (441mg, 4.694mmol) are added, so After be heated to 70 DEG C, keep 16h.Then reactant mixture is cooled to room temperature and be diluted with water.By reactant mixture acetic acid Ethyl ester extracts (3x50mL).The organic layer of merging is washed with water (2x50mL) and salt solution (50mL), through anhydrous Na₂SO₄Dry, subtract Pressure removes solvent, obtains crude residue.Crude mixture is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent： 90% ethyl acetate/hexane), obtain the fluoro- N- of the chloro- 8- of (4S) -7- (pyridin-3-yl) -3,4- dihydro-Isosorbide-5-Nitrae-endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamide (410mg, 1.231mmol, 0.805mmol, yield：34.6%), its For pale solid (TLC:Eluant, eluent：Net ethyl acetate, R_f:0.4)LCMS(m/z)333.9(M+H)⁺。

Synthesize the chloro- 8- of (4S) -7- fluoro- N- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides

Triethylamine (1.63mL, 11.737mmol) and triphosgene (696mg, 2.347mmol) are successively added into (4S) -7- Chloro- fluoro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of 8-(500mg,2.347mmol) In solution in THF (20mL), 30min is then stirred at room temperature and PA (441mg, 4.694mmol) is added, so After be heated to 70 DEG C, keep 16h.Then reactant mixture is cooled to room temperature, be diluted with water.Aqueous layer with ethyl acetate is extracted (3x50mL).The organic layer of merging is washed and salt solution (50mL) with water (2x50mL), through anhydrous Na₂SO₄Dry, then decompression is removed Solvent is removed, crude residue is obtained.Crude mixture is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：90% second Acetoacetic ester/hexane), obtain the fluoro- N- of the chloro- 8- of (4S) -7- (pyridine -2- bases) -3,4- dihydro-Isosorbide-5-Nitraes-endo-methylene group pyrido [2, 3-b] [1,4] diaza- 5 (2H)-formamides (350mg, 1.051mmol, 34.18% yield), it is pale solid (TLC:Eluant, eluent：100% ethyl acetate, R_f:0.4), LCMS (m/z) 334.1 (M+H)⁺。

Synthesize (4S) -7- chloro- N- (pyridazine -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine- 5 (2H)-formamides

30 DEG C into pyridazine -4- formic acid (0.5g, 4.03mmol) solution add diphenyl phosphate azides (1.308mL, 6.04mmol) and in solution of the DIPEA (2.111mL, 12.09mmol) in THF (10mL), it is stirred at 0 DEG C under a nitrogen Mix.Reactant mixture is stirred into 2h at 30 DEG C, (4S) -7- chloro- 2,3,4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-endo-methylene group pyrido is then added [2,3-b] [1,4] diaza(0.473g,2.417mmol).Reactant mixture is stirred into 6h at 90 DEG C.THF is depressurized and steamed Hair, then by residue diluted with water and is extracted into DCM.Organic layer water, salt water washing, it is then dried over sodium sulfate.Subtract Press evaporation solvent.Crude compound is by using ether and pentane (1:1) grinding purifying, obtain product (320mg, 0.91mmol, 23% yield), it is pale solid.LCMS(m/z)316.9[M+H]⁺。

Synthesize the chloro- 3,4- dihydros -1,4- bridges of (4S)-N- (3H- [1,2,3] triazol [4,5-d] pyrimidin-7-yl) -7- sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to the chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyridos [2,3-b] of (4S) -7- under nitrogen gas stirring [1,4] diazaTriphosgene is added in the solution of (500mg, 2.56mmol) in tetrahydrofuran (THF) (10mL) (758mg, 2.56mmol) and 30min is stirred at room temperature, then add triethylamine (0.356mL, 2.56mmol) and 3H- [1,2, 3] triazol [4,5-d] pyrimidine -7- amine (417mg, 3.07mmol).Reactant mixture is stirred into 16hr at 60 DEG C.Supervised by TLC Survey reaction (10% methanol/DCM).Reactant mixture 25ml water quenchings are gone out, and extracted with 25ml ethyl acetate.Organic layer is through sulphur Sour sodium is dried and is concentrated under reduced pressure, and obtains crude compound.Crude product is added into 100-200 silicagel columns and washed with 3%DCM/MeOH It is de-, obtain pure compound (4S)-N- (3H- [1,2,3] triazol [4,5-d] pyrimidin-7-yl) chloro- 3,4- dihydros-Isosorbide-5-Nitraes-of -7- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (200mg, 0.527mmol, 20.63% yield), It is pale solid, LCMS (m/z) 357.9 [M+H]⁺。

Synthesize 3- (pyridine -2- bases) -2H- pyridos [1,2-a] [1,3,5] triazine -2,4 (3H)-diketone

Room temperature into solution of the pyridine -2- formic acid (1g, 8.12mmol) of stirred under nitrogen in toluene (25mL) plus Enter diphenyl phosphate azide (2.235g, 8.12mmol) and TEA (1.132mL, 8.12mmol) and 30min is stirred at room temperature.This Reactant mixture is stirred into 2hr at 80 DEG C afterwards.Then, reactant mixture is cooled to room temperature and filtered, and solid is washed with toluene, Obtain compound 3- (pyridine -2- bases) -2H- pyridos [1,2-a] [1,3,5] triazine -2,4 (3H)-diketone (600mg, 2.352mmol, 29.0% yield), LCMS (m/z) 241.2 [M+H]⁺。

Synthesize (R) -4- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- amine

Added in seal pipe (R)-(2,2- dimethyl -1,3- dioxolane -4- bases) methanol (3.000g, 22.70mmol), the suspension of 4- chloropyridines -2- amine (1.459g, 11.35mmol) and sodium (0.522g, 22.70mmol).Will be anti- Mixture is answered to stir 16h at 140 DEG C.Then, reactant mixture is cooled to room temperature, be dissolved in MeOH, and poured into frozen water simultaneously Extracted with EtOAc.Organic phase aqueous salt solu-tion, it is then dried over sodium sulfate, filter and evaporate and obtain crude compound.Slightly Produced compounds are eluted through silica gel chromatography and with 2-3%MeOH/DCM, obtain pure compound (1.1g, 21%), LCMS (m/z)225.2[M+H]⁺。

Synthesize (S) -4- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- amine

Added in seal pipe (S)-(2,2- dimethyl -1,3- dioxolane -4- bases) methanol (3.000g, 22.70mmol), the suspension of 4- chloropyridines -2- amine (1.459g, 11.35mmol) and sodium (0.522g, 22.70mmol).Will be anti- Answer mixture to stir 16h at 140 DEG C, be subsequently cooled to room temperature, be dissolved in MeOH and pour into frozen water and extracted with EtOAc.It is organic Aqueous salt solu-tion is mutually used, it is then dried over sodium sulfate, filter and evaporate.Thick material uses 2-3% through silica gel chromatography MeOH/DCM is eluted, and obtains desired product (1.2g, 22%), LCMS (m/z) 225.2 [M+H]⁺。

Synthesize (R) -2- (tetrahydrofuran -3- bases epoxide) pyrimidine -4- amine

NaH is added into the solution of stirring of (R)-tetrahydrofuran -3- alcohol (2.72g, 30.9mmol) in THF (30mL) (0.926g, 23.16mmol), is then stirred at room temperature 30min.Be added portionwise thereto 2- chlorine pyrimidine -4- amine (2.0g, 15.44mmol), about 15min is lasted, then 16h is heated at 70 DEG C.So that reactant mixture reaches room temperature, 0 is then cooled to DEG C, it is quenched and is extracted with ethyl acetate (3x50ml) with icy water.The organic layer of merging is through anhydrous sodium sulfate drying, and filtering is simultaneously It is concentrated under reduced pressure, obtains crude compound.Crude product is through flash column chromatography (silica gel：100-200 mesh), obtain (R) -2- (four Hydrogen furans -3- bases epoxide) pyrimidine -4- amine (1.6g, 8.839mmol, 51.5% yield), it is pale solid.

Synthesize (R) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- amine

Under being stirred under a nitrogen at 0 DEG C to add in 6- chloropyrazine -2- amine (5g, 38.6mmol) sodium hydride (2.316g, 57.9mmol) with (R)-(2,2- dimethyl -1,3- dioxolane -4- bases) methanol (5.61g, 42.5mmol) in tetrahydrochysene furan The solution muttered in (THF) (50mL).Reactant mixture stirs 16h at 80 DEG C.Reactant mixture is quenched with icy water, Ran Houcui Get in ethyl acetate.Organic layer is through Na₂SO₄Dry and solvent is evaporated under reduced pressure, obtain crude product.Crude product adds to silicagel column and is used in combination DCM/MeOH is eluted.Merge the fraction containing product and reduction vaporization, obtain required product (2.8g, 11.9mmol, 31%), LCMS (m/z)225.9[M+H]⁺。

N31748-19

Synthesize (S) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- amine

By 6- chloropyrazine -2- amine (0.980g, 7.57mmol), (S)-(2,2- dimethyl -1,3- dioxolane -4- Base) methanol (2g, 15.13mmol) and sodium (0.348g, 15.13mmol) is added in seal pipe, then heats 16hr at 130 DEG C, Then reactant mixture is quenched and is extracted with ethyl acetate (5 × 50mL) with methanol and icy water (100mL).What is merged has Machine layer washed with water, saturated brine solution, through anhydrous sodium sulfate drying, filter and concentrate, obtain product (1g, 4.26mmol, 28.2% yield), LCMS (m/z) 265.1 [M+H]⁺。

Synthesize (S) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -4- amine

To (S) being stirred at room temperature under a nitrogen-(2,2- dimethyl -1,3- dioxolane -4- bases) methanol 2- chlorine is added dropwise in the suspension of (10.20g, 77mmol) and NaH (4.63g, 116mmol) in tetrahydrofuran (THF) (50mL) Pyrimidine -4- amine (5g, 38.6mmol), lasts 15min.Reactant mixture is stirred into 16hr at 70 DEG C.Then, by reactant mixture Use NaHCO₃The aqueous solution is quenched, and is then extracted with EtOAc, through Na₂SO₄Dry and evaporate.Crude product adds to silicagel column and uses 50% Hex/EtOAc is eluted.The fraction of collection is evaporated, desired product (3g, 11.84mmol, 30.7% yield) is obtained, it is ash White solid, LCMS (m/z) 226.2 [M+H]⁺。

Synthesize (R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -4- amine

Last solution of the 1min in room temperature to sodium hydride (0.817g, 34.1mmol) in tetrahydrofuran (THF) (30mL) Middle addition (R)-(2,2- dimethyl -1,3- dioxolane -4- bases) methanol (3g, 22.70mmol) is in THF (5mL) In solution, 15min is then stirred at room temperature, 2- chlorine pyrimidine -4- amine (2.059g, 15.89mmol) then is added dropwise in room temperature.Will be anti- Mixture is answered to stir 16h at 65 DEG C.Reactant mixture is poured into water and extracted (3 × 100mL) with EtOAc.Then it will merge Organic layer water, aqueous salt solu-tion, it is dried over sodium sulfate, and evaporated to obtain 4.0g crude compounds.Crude compound is through post Chromatogram purification, is eluted using 100-200 silica gel mesh and with 2-3%MeOH/DCM, obtain pure compound (2.5g, 10.42mmol, 46%), LCMS (m/z) 226.2 [M+H]⁺。

Synthesize (S) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- amine

To 2- chloropyridine -4- amine (1.459g, 11.35mmol), (S)-(2,2- dimethyl -1,3- dioxolane -4- Base) sodium (0.522g, 22.70mmol) is added in suspension in methanol (3.0g, 22.70mmol).By reactant mixture 140 DEG C stirring 16hr, monitor reaction process.

Reactant mixture is dissolved in MeOH, pours into frozen water and is extracted (3 × 100mL) with EtOAc.Then having merging Machine layer water, aqueous salt solu-tion, it is dried over sodium sulfate, and evaporated to obtain 4.0g crude compounds.Crude compound is through column chromatography Purifying, is eluted using 100-200 silica gel mesh and with 2-3%MeOH/DCM, obtains (S) -2- ((2,2- dimethyl -1,3- dioxas Pentamethylene -4- bases) methoxyl group) pyridine -4- amine (2.5g, 10.73mmol, 47.3% yield), LCMS (m/z) 225.3 [M+H]⁺。

Synthesize (R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- amine

2- chloropyridine -4- amine (4g, 31.1mmol), (R)-(2,2- dimethyl -1,3- bis- are added in seal pipe in room temperature Tetrahydrofuran -4- bases) methanol (2.056g, 15.56mmol) and sodium (0.715g, 31.1mmol) solution.By reaction mixing Thing stirs 48hr at 140 DEG C.Reactant mixture is cooled to room temperature and priority MeOH and water quenching are gone out.Then reactive material is used EtOAc is extracted.Then organic layer priority water and aqueous salt solu-tion and sodium sulphate drying and dehydrating is used, filters and be completely distilled off. Crude product adds to silicagel column and with Hex/EtOAc (1:1) elute.The fraction of collection is evaporated, desired product is obtained (2.250g, 9.93mmol, 31.9% yield), LCMS (m/z) 225.0 [M+H]⁺。

Synthesize (S) -2- ((tetrahydrofuran -3- bases) epoxide) pyrimidine -4- amine

In the molten of stirring of the room temperature to 2- chlorine pyrimidine -4- amine (2g, 15.44mmol) in tetrahydrofuran (THF) (20mL) NaH (0.741g, 30.9mmol) is added dropwise in liquid, 5min time is lasted.Then by reactant mixture in 30 DEG C of stir abouts 10min.(S)-tetrahydrofuran -3- alcohol (1.088g, 12.35mmol), Ran Hou are added into above-mentioned reactant mixture at 30 DEG C 80 DEG C are stirred 8 hours.Reactant mixture icy water is quenched and is extracted with ethyl acetate at 0 DEG C.Organic layer is abundant with water Washing, and through Na₂SO₄Dry.Solvent is evaporated under reduced pressure, product is obtained.Crude product is ground with petroleum ether, [the M+ of LCMS (m/z) 182.2 H]⁺。

Synthesize (1- methyl isophthalic acid H- pyrazoles -4- bases) phenyl carbamates

Phenyl chloroformate (2.90g, 18.53mmol) is added into pyridine (3.12mL, 38.6mmol) in dichloromethane at 0 DEG C (DCM) in the solution of the stirring in (50mL), then stir 15min, then identical temperature add 1- methyl isophthalic acid H- pyrazoles- 4- amine (1.5g, 15.45mmol).4h is stirred at room temperature in reactant mixture.After initiation material exhausts (being monitored through TLC), add Icy water, the organic layer water and salt water washing of separation.Organic layer is filtered through sodium sulphate and concentrated, and obtains rough chemical combination Thing.Crude compound is purified through column chromatography, is eluted, must be expired using 60-120 (silica gel) and in 50% ethyl acetate/hexane The product (1.6g, 6.41mmol, 42% yield) of prestige, it is light tan solid, LCMS (m/z) 218.1 (M+H)⁺。

Pyridine synthesis -3- aminocarbamic acid phenyl esters

Under nitrogen gas stirring room temperature to phenyl chloroformate (2.163g, 13.81mmol) and pyridine (1.375mL, Pyridine -3- amine (1.0g, 10.63mmol) 17.00mmol) is added in the solution in dichloromethane (DCM) (30mL).Will reaction 30min is stirred at room temperature in mixture.Reactant mixture is quenched with saturated sodium bicarbonate solution.Organic layer is separated, water layer is used DCM (50mL) is extracted.The DCM layers of merging are washed with water and use sodium sulphate drying and dehydrating, filter and high vacuum concentration, are slightly produced Thing.Crude product adds to silicagel column and uses 20%EtOAc/ Hex.The fraction of collection is evaporated, desired product is obtained (1.3g, 6.01mmol, 57%), it is white solid, LCMS (m/z) 215.1 (M+H)⁺。

Synthesize pyrimidine -2 --amino phenyl formate

Under a nitrogen phenyl chloroformate (2.140g, 13.67mmol) from room temperature to stirring and pyridine (1.361mL, Pyrimidine -2- amine (1.0g, 10.51mmol) 16.82mmol) is added in the solution in dichloromethane (DCM) (10mL).Will reaction 30min is stirred at room temperature in mixture.Reactant mixture is quenched with saturated sodium bicarbonate solution.Organic layer is separated, water layer is used DCM (50mL) is extracted.The DCM layers of merging are washed with water and use sodium sulphate drying and dehydrating, filter and high vacuum concentration, are slightly produced Thing.Added to silicagel column and use 20%EtOAc/ Hex.The fraction of collection is evaporated, obtain desired product (1.6g, 6.49mmol, 61.7%), LCMS (m/z) 216.3 (M+H)⁺。

Synthesize (5- fluorine pyridine -2- bases) phenyl carbamates

Under a nitrogen phenyl chloroformate (1.397g, 8.92mmol) from room temperature to stirring and pyridine (0.721mL, 5- fluorine pyridine -2- amine (1.0g, 8.92mmol) 8.92mmol) is added in the solution in dichloromethane (DCM) (40mL).Will be anti- Answer mixture that 30min is stirred at room temperature.Reactant mixture is quenched with saturated sodium bicarbonate solution.Organic layer is separated, by water layer Extracted with DCM (20mL).The organic layer priority water and aqueous salt solu-tion of merging simultaneously use sodium sulphate drying and dehydrating, filter and true Sky concentration, obtains desired product (1.4g, 5.94mmol, 67%), LCMS (m/z) 233.2 (M+H)⁺。

Synthesize (2- methyl -2H- indazole -5- bases) phenyl carbamates

Under nitrogen gas stirring room temperature to phenyl chloroformate (1.064g, 6.79mmol) and pyridine (0.550mL, 2- methyl -2H- indazole -5- amine (1g, 6.79mmol) 6.79mmol) is added in the solution in dichloromethane (DCM) (40mL). 30min is stirred at room temperature in reactant mixture.Reactant mixture is quenched with saturated sodium bicarbonate solution.Separate organic layer, water Layer is extracted (20mL) with DCM.The organic layer priority water and aqueous salt solu-tion of merging and with sodium sulphate drying and dehydrating and vacuum Concentration, obtains (2- methyl -2H- indazole -5- bases) phenyl carbamates (1.3g, 4.82mmol, 70.9% yield), LCMS (m/z)268.1(M+H)⁺。

Synthesize (5- ethyl pyrazine -2- bases) phenyl carbamates

Pyridine (1.051mL, 12.99mmol) is added dropwise into phenyl chloroformate (1.324mL, 10.56mmol) in room temperature to exist In the solution of stirring in dichloromethane (DCM) (20ml) and stirring 30 minutes.Then 5- ethyl pyrazine -2- amine is added dropwise in room temperature The solution of (1g, 8.12mmol) in dichloromethane (DCM) (10ml), then stirs 16h at 50 DEG C.Reach reactant mixture Room temperature, is diluted with DCM (3X50mL), is washed with water (2X30mL) and salt solution (30mL).Organic layer is separated, it is then dry through sodium sulphate It is dry, filter and concentrate.Residue is purified through column chromatography, using silica gel (100-200 mesh), is made with 10% ethyl acetate/petroleum ether For eluant, eluent, required product is obtained, it is canescence fluffy solid (1.6g, 6.58mmol, 81%), (M of LCMS (m/z) 244.2 +H)⁺。

Synthesize (5- cyclopropyl pyrazine -2- bases) phenyl carbamates

Pyridine (0.598mL, 7.40mmol) is added dropwise to phenyl chloroformate (0.928mL, 7.40mmol) in dichloro at 0 DEG C In the solution of stirring in methane (DCM) (15ml), then it is stirred at room temperature 30 minutes.Then 5- cyclopropyl pyrroles are added dropwise at 0 DEG C Solution (DCM) (5ml) of the piperazine -2- amine (1g, 7.40mmol) in dichloromethane, is then stirred at room temperature 3h.By reaction mixing Thing DCM dilutes (3X50mL), is washed with water (2X20mL) and salt solution (20mL).Organic layer is separated, it is then dried over sodium sulfate, Filter and concentrate.Residue is purified through column chromatography, using silica gel (100-200 mesh), using 10% ethyl acetate/petroleum ether as washing De- agent, obtains desired product (1.4g, 5.31mmol, 72%), and it is canescence the fluffy solid, (M+ of LCMS (m/z) 256.2 H)⁺。

Synthesize (6- ethyoxyl pyrazine -2- bases) phenyl carbamates

Pyridine (0.930mL, 11.50mmol) is added into phenyl chloroformate (1.463g, 9.34mmol) in DCM in room temperature In solution in (15mL) and stirring 20min, then add 6- ethyoxyl pyrazine -2- amine (1.0g, 7.19mmol) in DCM Solution in (15mL), is then further continued for stirring 40min.Reactant mixture is diluted into (2X20mL) with DCM, with water (20mL × 2) washed with salt solution (10mL).Organic extract is through Na₂SO₄Dry, solvent is then removed in vacuum, obtain desired product (1.65g, 5.22mmol, 72.6% yield), it is yellow solid, LCMS (m/z) 260.2 (M+H)⁺。

Synthesize pyridazine -3- aminocarbamic acid phenyl esters

At 25 DEG C to phenyl chloroformate (1.070g, 6.83mmol), pyridine (0.665g, 8.41mmol) under nitrogen gas stirring Suspension of the pyridazine -3- amine (0.5g, 5.26mmol) in dichloromethane (5ml) is added in solution in dichloromethane (10ml) Liquid, continues 5min.Reactant mixture is stirred into 1hr. then at 25 DEG C, organic phase is washed with 3mL water, 3mL saturated brines, through sulphur Sour sodium is dried and is concentrated in vacuo, and obtains crude product, it is white solid.Compound is washed with hexane, is dried under reduced pressure, LCMS (m/ z)216.2(M+H)⁺。

Synthesize pyrimidine-4-yl phenyl carbamate

At 25 DEG C to phenyl chloroformate (1.070g, 6.83mmol), pyridine (0.665g, 8.41mmol) under nitrogen gas stirring It is added dropwise in suspension of the pyrimidine -4- amine (0.5g, 5.26mmol) in DCM (5ml), continues in solution in DCM (15ml) 5min.Reactant mixture is stirred into 1hr at 25 DEG C.Organic phase 3mL water, 3mL salt water washings, it is dried over sodium sulfate and depressurize dense Contracting, obtains crude product, it is pale solid.Crude compound is washed with hexane, is then dried under reduced pressure, and obtains desired product (500mg, 1.95mmol, 37%), LCMS (m/z) 215.9 (M+H)⁺。

The senior bicyclic intermediate of synthesis

Synthesize (4S)-N- (4- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- Base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide

In room temperature to (4R) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- bridge methylenes under nitrogen gas stirring Yl pyridines simultaneously [2,3-b] [1,4] diazaThe solution of (499.5mg, 1.636mmol) in tetrahydrofuran (THF) (20mL) Middle addition TEA (1.368mL, 9.82mmol), triphosgene (486mg, 1.636mmol).15min is stirred at room temperature, then Last 5min be added dropwise (R) -4- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- amine (1101mg, 4.91mmol) the solution in THF (5mL).Reactant mixture is stirred into 16h at 65 DEG C, is subsequently poured into water and is extracted with EtOAc Take (3 × 100mL).Then it is dried over sodium sulfate, and evaporated by organic layer water, the aqueous salt solu-tion of merging, slightly produced Thing.The inverted chromatogram of crude compound (0.1%HCOOH＆ water)/MeOH is purified, and obtains desired product (450mg, 49%), LCMS(m/z)555.9(M+H)⁺。

Synthesize (4S)-N- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- Base) -7- (6- picoline -3- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide

At 30 DEG C to (R) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- amine Triphosgene (423mg, 1.427mmol) is added in the solution of (803mg, 3.57mmol) in tetrahydrofuran (THF) (20mL), so Afterwards 30min is stirred in identical temperature.Then TEA (1.657mL, 11.89mmol) and (4S) -7- (6- is successively added in room temperature Picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(600mg, 2.378mmol).Reactant mixture is stirred into 16hr at 65 DEG C.Pass through TLC and thick LCMS monitoring reactions.THF is evaporated under reduced pressure, Residue diluted with water, is then extracted into DCM.Organic layer is through Na₂SO₄Dry, solvent is evaporated under reduced pressure, crude product is obtained.It is rough Compound is purified through column chromatography, and eluant, eluent is used as using silica gel as stationary phase (100-200 mesh) and 2-3%MeOH/EtOAc. By pure fraction collector and be concentrated under reduced pressure, obtain pure product 0.4g, its be pale solid (450mg, 0.71mmol, 30%), LCMS (m/z) 504.3 (M+H)⁺。

Synthesize (4S)-N- (4- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- Base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide

In room temperature to (4R) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- bridge methylenes under nitrogen gas stirring Yl pyridines simultaneously [2,3-b] [1,4] diazaIn the solution of (545mg, 1.785mmol) in tetrahydrofuran (THF) (20mL) Add TEA (1.493mL, 10.71mmol), triphosgene (530mg, 1.785mmol).Then reactant mixture is stirred into 15min, Then last 5min and (S) -4- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- amine is added dropwise The solution of (1201mg, 5.36mmol) in THF (5mL).Reactant mixture is stirred into 16h at 65 DEG C, room temperature is subsequently cooled to And be poured into water and extracted (3 × 100mL) with EtOAc.Then by organic layer water, the aqueous salt solu-tion of merging, through sulfuric acid Sodium is dried and evaporated, and obtains crude product.By its inverted column chromatography purifying (0.1%HCOOH＆ water)/MeOH, desired production is obtained Thing (500mg, 49%), LCMS (m/z) 556.3 (M+H)⁺。

Synthesize (4S)-N- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- Base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide

25 DEG C to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diazaThree light are added in the solution of the stirring of (850mg, 2.78mmol) in 40mL THF (seal pipe) Gas (324mg, 1.093mmol) and stirring 30min.(R) -6- ((2,2- dimethyl -1,3- two are added into the reactant mixture Tetrahydrofuran -4- bases) methoxyl group) pyrazine -2- amine (938mg, 4.16mmol), then by reactant mixture in 65 DEG C of stirrings 16h.So that reactant mixture is cooled to room temperature and poured the mixture into cold water (70mL), and it is extracted with ethyl acetate (3x50mL).The organic layer of merging is through anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain crude product.Crude mixture is through fast Fast column chromatography purifies (silica gel；100-200 mesh, with 1 to 2% ethanol/methylene elute), obtain (4S)-N- (6- (((R) -2, 2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- two Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(900mg, 1.618mmol 57% is received -5 (2H)-formamides Rate), it is white solid (TLC:Eluant, eluent：10%MeOH/DCM, R_f=0.3), LCMS (m/z) 557.3 (M+H)⁺。

Synthesize (4S)-N- (6- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- Base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide

To (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diazaIn the solution of (723mg, 2.368mmol) in tetrahydrofuran (THF) (25mL) add triphosgene (351mg, 1.184mmol), 30min then is stirred at room temperature in reactant mixture, be subsequently added into TEA (1.650mL, 11.84mmol) and (S) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- amine (800mg, 3.55mmol).Will Reactant mixture stirs 16hr at 65 DEG C, is subsequently cooled to room temperature and is poured into water and is extracted (3 × 100mL) with EtOAc.Then It is dried over sodium sulfate, and evaporated by organic layer water, the aqueous salt solu-tion of merging, crude compound is obtained, its is inverted Post is purified and eluted with 83% (0.1%HCOOH＆ water)/MeOH, obtains pure compound (300mg, 0.534mmol, 23%), LCMS(m/z)557.4(M+H)⁺。

Synthesize (4S)-N- (2- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -4- Base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide

To (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diazaIn the solution of (600mg, 1.965mmol) in tetrahydrofuran (THF) (10mL) add triphosgene (583mg, 1.965mmol), TEA (1.644mL, 11.79mmol), is stirred at room temperature 15min, then add (S) -2- ((2,2- dimethyl - 1,3- dioxolane -4- bases) methoxyl group) solution of the pyrimidine -4- amine (1328mg, 5.90mmol) in THF (5mL), go through When 1min.Reactant mixture is stirred into 16hr at 65 DEG C, is subsequently poured into water and is extracted (3 × 50mL) with EtOAc.Then it will close And organic layer water, aqueous salt solu-tion, it is dried over sodium sulfate, and evaporated, obtain 600mg.The inverted post of crude compound is pure Change and eluted with 90% (0.1%HCOOH＆ water)/MeOH, obtain desired compound (450mg, 0.76mmol, 39%), LCMS (m/z)557.2(M+H)⁺。

Synthesize (4S)-N- (2- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- Base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide

To (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diazaIn the solution of (600mg, 1.965mmol) in tetrahydrofuran (THF) (10mL) add TEA (1.644mL, 11.79mmol), triphosgene (583mg, 1.965mmol), stirs 15min and (S) -2- ((2,2- dimethyl -1,3- dioxies is added dropwise Heterocycle pentane -4- bases) methoxyl group) pyridine -4- amine (1322mg, 5.90mmol).Reactant mixture is stirred into 16hr at 65 DEG C, and Reaction process is monitored by TLC.Reactant mixture is poured into frozen water and extracted (3 × 100mL) with EtOAc.Then it will merge Organic layer water, aqueous salt solu-tion, it is dried over sodium sulfate, and evaporated, obtain crude product.The inverted post of crude compound is pure Change and eluted with 93% (0.1%HCOOH＆ water)/MeOH, obtaining desired product, (450mg, 0.807mmol, 41.1% are received Rate), LCMS (m/z) 556.4 (M+H)⁺。

To (4R) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diazaIn the solution of (400mg, 1.310mmol) in tetrahydrofuran (THF) (10mL) add TEA (1.096mL, 7.86mmol), triphosgene (389mg, 1.310mmol), is stirred at room temperature 15min.Add (R) -2- ((2,2- dimethyl -1,3- Dioxolane -4- bases) methoxyl group) solution of the pyrimidine -4- amine (885mg, 3.93mmol) in THF (2.0mL).Will reaction Mixture stirs 16hr at 65 DEG C.Reactant mixture is poured into water and extracted (3 × 100mL) with EtOAc.Then by merging Organic layer water, aqueous salt solu-tion, it is dried over sodium sulfate, and evaporated to dry.The purifying of crude compound inverted post and use 90% (0.1%HCOOH＆ water)/MeOH is eluted, and obtains pure compound (350mg, 0.602mmol, 46%), LCMS (m/z) 557.0 (M+H)⁺。

Synthesize (4S)-N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- Base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide

In room temperature to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- bridge methylenes under nitrogen gas stirring Yl pyridines simultaneously [2,3-b] [1,4] diazaAdded in the solution of (1g, 3.28mmol) in tetrahydrofuran (THF) (15mL) TEA (2.74mL) and triphosgene solution, are stirred 30 minutes.(R) -2- ((2,2- dimethyl -1,3- dioxas are added thereto Pentamethylene -4- bases) methoxyl group) pyridine -4- amine (2.204g, 9.83mmol) in tetrahydrofuran (THF) (8mL) (2.204g) plus Enter.Reactant mixture is stirred into 16hr at 60 DEG C.Reactant mixture is concentrated, residue is absorbed in DCM (100mL).Will Solution water and salt water washing, through Na₂SO₄It is dried, filtered and concentrated.Crude product adds to silicagel column and uses EtOAc/ petroleum ethers (60:20) elute, the fraction of collection is evaporated, desired product (600mg, 0.92mmol, 28%) is obtained, it is canescence half Solid, LCMS (m/z) 556.3 (M+H)⁺。

Synthesize (4S) -7- (2- picoline -4- bases)-N- (2- (pyridin-3-yl) ethyl) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

It is sub- to (4S) -7- (2- picoline -4- the bases) -2,3,4,5- tetrahydrochysene -1,4- bridges being stirred at room temperature under a nitrogen Picoline simultaneously [2,3-b] [1,4] diaza(400mg, 1.585mmol), triethylamine (1.105mL, 7.93mmol) and three 2- (pyridin-3-yl) ethamine is added in solution of the phosgene (282mg, 0.951mmol) in tetrahydrofuran (THF) (20mL) The solution of (387mg, 3.17mmol) in THF (5mL).Reactant mixture is stirred into 16hr at 65 DEG C, and monitored instead by TLC Answer process.Reactant mixture is poured into frozen water and extracted (3 × 100mL) with EtOAc.Then by the organic layer water of merging, Aqueous salt solu-tion, it is dried over sodium sulfate, and evaporated, obtain crude compound.The inverted post purifying of crude compound and 25- 30% (0.1%HCOOH＆ water)/MeOH is eluted, and obtains final product (250mg, 0.599mmol, 37.8% yield), LCMS (m/ z)401.1(M+H)⁺。

Synthesize 4- ((4S) -5- (pyrazine -2- bases carbamoyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 7- bases) piperazine -1- t-butyl formates

20 DEG C to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (600mg, 1.894mmol) and piperazine -1- t-butyl formates (706mg, 3.79mmol) exist Cs is sequentially added in the solution of degassing in 1,4- dioxanes (10mL)₂CO₃(1852mg, 5.68mmol), xphos (361mg, 0.758mmol) and PdOAc₂(85mg,0.379mmol).Reactant mixture is stirred into 16hr at 100 DEG C.Reactant mixture is fallen Enter in cold water (20mL) and be extracted with ethyl acetate (50mL).Organic layer is through anhydrous sodium sulfate drying and is concentrated under reduced pressure, and obtains thick Product.Crude product adds to silicagel column and eluted with 3%DCM/MeOH, obtain 4- ((4S) -5- (pyrazine -2- bases carbamoyl) - 2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 7- bases) piperazine -1- t-butyl formates (359mg, 0.616mmol, 32.5% yield), LCMS (m/z) 467.3 (M+H)⁺。

Synthesize 3- methyl -4- ((4S) -5- (pyrazine -2- bases carbamoyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 7- bases) piperazine -1- t-butyl formates

20 DEG C to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (800mg, 2.53mmol), 3- methyl piperazine -1- t-butyl formates (1012mg, 5.05mmol) Cs is sequentially added in the solution of the degassing in 1,4- dioxanes (10mL)₂CO₃(2469mg, 7.58mmol) and Xphos (482mg, 1.010mmol), PdOAc₂(113mg,0.505mmol).Reactant mixture is stirred into 16hr at 100 DEG C.It is logical Cross TLC monitoring reactions.Reactant mixture is poured into cold water (20mL) and is extracted with ethyl acetate (50mL).Organic layer is through anhydrous Sodium sulphate is dried, and is concentrated under reduced pressure, is obtained crude product.Crude product adds to silicagel column and eluted with 2%DCM/MeOH.By the level of collection Divide evaporation, obtain desired product (397.5mg, 0.670mmol, 26.5% yield), LCMS (m/z) 481.1 (M+H)⁺。

Synthesize (4S) -7- (4- benzyl -3- methylpiperazine-1-yls)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

20 DEG C to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (500mg, 1.579mmol), 1- benzyl -2- methyl piperazines (601mg, 3.16mmol) exist Cs is sequentially added in the solution of degassing in 1,4- dioxanes (20mL)₂CO₃(1543mg, 4.74mmol) and cinnamoyl chlorine [1, 3- bis- (diisopropyl phenyl) -2- imidazolidinyls subunit] Pd (II) (51.3mg, 0.079mmol).By reactant mixture 110 DEG C stirring 16 hours.Reactant mixture is poured into cold water (50mL) and is extracted with ethyl acetate (200mL).Organic layer is through anhydrous sulphur Sour sodium is dried and is concentrated under reduced pressure, and obtains crude product.Crude product adds to silicagel column and eluted with 3%DCM/MeOH, obtains (4S) -7- (4- benzyl -3- methylpiperazine-1-yls)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (501.5mg, 0.885mmol, 56.0% yield), LCMS (m/z) 471.3 (M+H)⁺。

Synthesize 4- bromo- 2- (difluoromethyl) pyridine

DAST (0.620mL, 4.69mmol) is added dropwise into 4- bromopyridine -2- formaldehyde (700mg, 3.76mmol) at 0 DEG C to exist In solution in chloroform (21mL).Reactant mixture is stirred into 12h at 20 DEG C.Reactant mixture is poured into saturation NaHCO₃Solution In (20mL), and extracted (2X20mL) with DCM.DCM layers through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain 4- bromo- 2- (difluoromethyl) pyridine (400mg, 1.870mmol, 49.7% yield), it is the light yellow solid, [M+ of LCMS (m/z) 208.0 H]⁺。

Synthesize (2- (difluoromethyl) pyridin-4-yl) boric acid

28 DEG C by potassium acetate (472mg, 4.81mmol) add to 4- bromo- 2- (difluoromethyl) pyridine (400mg, 1.923mmol), and double solution of (pinacol conjunction) diborane (610mg, 2.404mmol) in the dioxane of Isosorbide-5-Nitrae-(10mL).Will Reactant mixture degassing 15min, adds PdCl₂(dppf)(4.22mg,5.77μmol).Reactant mixture is deaerated 15min again, And reactant mixture is stirred into 48hr at 80 DEG C.Reactant mixture is cooled to 28 DEG C, evaporation, by crude product distribution in water Between (10mL) and EtOAc (25mL).EtOAc layers of separation, then through anhydrous Na₂SO₄Dry, filtering, and filtrate is evaporated, obtain To (2- (difluoromethyl) pyridin-4-yl) boric acid (330mg, 1.107mmol, 57.6% yield), it is brown solid, LCMS (m/z)174.1[M+H]⁺。

Synthesize 4- benzyl -3- methyl piperazine -1- t-butyl formates

0 DEG C to 3- methyl piperazine -1- t-butyl formates (1g, 4.99mmol) at N,N-dimethylformamide (DMF) K is added in solution in (100mL)₂CO₃(2.070g,14.98mmol).After 0 DEG C is stirred 10min, benzyl bromide a-bromotoluene is added dropwise (0.891mL, 7.49mmol) and reactant mixture is stirred into 16hr at 35 DEG C and reaction is monitored by TLC.By reactant mixture Pour into NaHCO₃In the aqueous solution (50mL) and it is extracted with ethyl acetate (200mL), with priority water and salt water washing.Organic layer is through nothing Aqueous sodium persulfate is dried, and is concentrated under reduced pressure, is obtained crude product.Crude product adds to silicagel column and eluted with 10%Hex/EtOAc.It will collect Fraction evaporation, obtain 4- benzyl -3- methyl piperazine -1- t-butyl formates (1g, 3.17mmol, 63.4% yield), LCMS (m/ z)174.1[M+H]⁺。

Synthesize 1- benzyl -2- methyl piperazines

0 DEG C to 4- benzyl -3- methyl piperazine -1- t-butyl formates (1.4g, 4.82mmol) at dichloromethane (DCM) TFA (1.857mL, 24.10mmol) is added in solution in (25mL) and reactant mixture is stirred into 3hr at 35 DEG C.Pass through TLC Monitoring reaction.Solvent is evaporated under reduced pressure, crude product is obtained.By residue with triturated under ether (2 × 50mL).By the filtering of gained solid simultaneously Washed with ether.It is dried under reduced pressure, obtains 1- benzyl -2- methyl piperazine trifluoroacetates (800mg, 2.63mmol, 54.5% Yield)

Synthesize ((R) -2- methyl morpholine generations) (4S) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-yls) ketone:

Triphosgene (0.529g, 1.783mmol) is added into triethylamine (1.243mL, 8.92mmol) and (4S) -7- in room temperature (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(0.45g, 1.783mmol) in the solution of the stirring in tetrahydrofuran (50mL) and stirring 1h, be subsequently added into (R) -2- methyl morpholine hydrochloric acid Salt (0.368g, 2.68mmol), is then heated to 70 DEG C, keeps 15h.Be cooled to room temperature, then with ethyl acetate (100mL) and Water (100mL) dilutes.The organic layer water and salt water washing of separation.Organic layer is dried over sodium sulfate, filters and is concentrated under reduced pressure, obtains To crude compound (TLC eluant, eluents：10%MeOH/ ethyl acetate；UV is activated；R_f~0.4).Crude compound passes through column chromatography Purifying, is eluted using neutral alumina and in 50% ethyl acetate/hexane, obtains (R) -2- methyl morpholine generations) ((4S) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-yls) first Ketone (0.2g, 0.514mmol, 32.4% yield), it is colloidal cpd, LCMS (m/z) 380.3 (M+H)⁺。

Synthesize ((S) -2- methyl morpholine generations) ((4S) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-yls) ketone

Triphosgene (0.470g, 1.585mmol) is added into triethylamine (1.105mL, 7.93mmol) and (4S) -7- in room temperature (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(0.4g, 1.585mmol) in the solution of the stirring in tetrahydrofuran (50mL) and stirring 1h, be subsequently added into (S) -2- methyl morpholine hydrochloric acid Salt (0.327g, 2.378mmol), is then heated to 70 DEG C, keeps 15h.It is cooled to room temperature and with ethyl acetate (100mL) and water (100mL) dilutes.The organic layer water and salt water washing of separation.Organic layer is dried over sodium sulfate, filters and is concentrated under reduced pressure, obtains Crude compound (TLC eluant, eluents：10%MeOH/ ethyl acetate；UV is activated；R_f~0.4).Crude compound is pure by column chromatography Change, eluted using aluminum oxide and in 50% ethyl acetate/hexane, obtain (S) -2- methyl morpholine generations) ((4S) -7- (2- methyl Pyridin-4-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-yls) ketone (0.2g, 0.514mmol, 32.4% yield), it is colloidal cpd, LCMS (m/z) 480.3 (M+H)⁺。

Synthesis (4S) -8- chloro- 7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza

At 0 DEG C to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- bridge methylenes under nitrogen gas stirring Yl pyridines simultaneously [2,3-b] [1,4] diazaN- chlorine ambers are added dropwise in the solution of (10.0g, 32.8mmol) in chloroform (50mL) Amber acid imide (6.56g, 49.1mmol), continues 10min.Reactant mixture is stirred into 6hr at 50 DEG C., will be mixed after reaction terminates Compound is quenched with icy water (60mL) and extracted (3x60ml) with DCM, is separated DCM layers and with salt water washing (2x30ml), is divided From DCM layers and through anhydrous sodium sulfate drying, filtered and concentrated, obtain thick material.Thick material is quick through neutral alumina Chromatogram purification.Thick material is diluted with DCM, then absorbed and with 30%EtOAc/ petroleum ethers -50% with neutral alumina EtOAc/ petroleum ethers are eluted.Collect fraction and concentrate, obtain the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetra- Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(6.3g, 18.12mmol, 55.3% yield), it is yellowish Color solid, LCMS (m/z) 340.1 [M+H]⁺。

Synthesize (4S) -8- chloro- N- (5- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -3- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

At 0 DEG C by the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza(500mg, 1.472mmol), triethylamine (1.231mL, 8.83mmol) is absorbed in tetrahydrofuran (THF) in (50mL), gained yellow solution is stirred into 10min.Then 0 DEG C it is disposable add triphosgene (437mg, 1.472mmol).45min is stirred at room temperature in gained yellow suspension at 0 DEG C.At 0 DEG C, by (S) -5- ((2,2- dimethyl -1, 3- dioxolane -4- bases) methoxyl group) THF (4mL) solution of pyridine -3- amine (330mg, 1.472mmol) adds to above-mentioned Huang Color contamination suspension, lasts 5min time.Gained yellow suspension is heated to 70 DEG C, 24hr is kept.Reaction process is through TLC10% MeOH/DCM is monitored, and TLC forms multiple spots after indicating 24h.Reactive material is cooled to room temperature, with water (20mL), acetic acid second Ester (30mL*2) dilutes.The organic layer of merging is concentrated under reduced pressure with salt water washing (15mL), filtering dried over sodium sulfate, is obtained palm fibre Color solid.Crude product is purified through combiflash chromatograms on the silica gel of 230-400 mesh sizes.By post MeOH/DCM gradient Elution.Desired compound is eluted with 7%MeOH/DCM.Fraction containing pure compound is concentrated under reduced pressure, (4S) -8- is obtained Chloro- N- (5- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridin-3-yl) -7- (3- (fluoroforms Base) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (210mg, 0.331mmol, 22.47% yield), it is pale solid, LCMS (m/z):590.15[M+H]⁺。

To the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaTriethylamine is added in the solution of (7g, 20.60mmol) in tetrahydrofuran (THF) (120mL) (17.23mL, 124mmol) and triphosgene (6.11g, 20.60mmol).30min is stirred at room temperature in reactant mixture.It is anti-to this Answer addition (R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -4- amine in mixture (11.60g, 51.5mmol) and stir 12hr at 65 DEG C.Reactant mixture is cooled to room temperature, evaporating completely solvent under decompression, And distribute between water (100mL) and EtOAc (2 × 200mL).Organic layer is separated, through anhydrous Na₂SO₄Dry, filter and will filter Liquid evaporates, and obtains crude product.Purified on column chromatography is purified, using neutral alumina and with 25-30%EtOAc/ hexane (gradients System) elution, desired product (7.2g) is obtained, it is white solid.(100mL) dilutes product (6.9g) in ethanol, so Handled afterwards with Silicycle palladiums scavenger (3.5g), then stir 3hr at 55 DEG C.Reactant mixture is passed through into Celite pad mistake Filter, and Celite pad is washed with hot ethanol (50ml), the filtrate decompression of acquisition is concentrated, the chloro- N- (2- of (4S) -8- are obtained (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine-4-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (6.9g, 11.65mmol, 56.6% yield), it is white solid.LCMS(m/z):591.16[M+H]⁺。

Synthesize (4S) -8- chloro- N- (4- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

Under nitrogen gas stirring room temperature to the chloro- 7- of solid (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysenes - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(3.0g, 8.83mmol) is in tetrahydrofuran (THF) (30mL) Solution in add Solid triphosgene (1.572g, 5.30mmol), be stirred at room temperature under a nitrogen 30 minutes.Then add thereto Enter DIPEA (7.71mL, 44.2mmol) and (S) -4- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrrole Pyridine -2- amine (2.97g, 13.25mmol), 16h is kept under the conditions of seal pipe at 75 DEG C.Pass through TLC and LCMS monitoring reactions.Will Reactant mixture is concentrated, and residue is absorbed in into dichloromethane (100mL).By solution water and salt water washing, through Na₂SO₄It is dry It is dry, filter and concentrate, obtain crude compound.Crude product is added in neutral alumina, and is eluted with 50%EtOAc/ petroleum ethers. Collect fraction and be concentrated to give compound, washed with pentane, obtain pure compound, LCMS (m/z) 590.43 [M+H]⁺。

Synthesize (4S) -8- chloro- N- (5- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Piperazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

In room temperature to the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diazaIn the solution of the stirring of (500mg, 1.472mmol) in tetrahydrofuran (THF) (20mL) Triphosgene (437mg, 1.472mmol) is added, triethylamine (1.231mL, 8.83mmol) is subsequently added into.Reactant mixture is stirred 45min, then adds (R) -5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- amine The solution of (663mg, 2.94mmol) in tetrahydrofuran (THF) (5mL).Reactant mixture is stirred into 12hr at 65 DEG C.TLC refers to Show that initiation material exhausts and forms new spot.Water (25mL) is added into reactant mixture.Water layer is extracted with EtOAc (2 × 25mL), the organic layer of merging is through anhydrous Na₂SO₄It is dried, filtered and concentrated, obtains desired crude product.Purified on column chromatography Purifying, using 100-200 silica gel (eluant, eluent 35-50%EtOAc/ petroleum ethers), obtains desired pure product (4S) -8- chloro- N- (5- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- bases) -7- (3- (trifluoromethyl) Phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (500mg, 0.837mmol, 56.9% yield), it is pale solid, LCMS (m/z):590.8[M+H]⁺。

Synthesize (4S) -8- chloro- N- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Piperazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

DIPEA (13.69g, 106mmol) and triphosgene (10.48g, 35.3mmol) are added into (4S) -8- in 25 DEG C of priorities Chloro- 7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza In the solution of (12.0g, 35.3mmol) in tetrahydrofuran (THF) (200mL), stir 1h and add (R) -6- ((2,2- diformazans Base -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- amine (15.91g, 70.6mmol) and 70 DEG C heat 18h.Will Reactant mixture is cooled to 28 DEG C, and distributes between water (50mL) and EtOAc (100mL).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into crude product.Crude product purifies (post through combiflash column chromatographys：C18, is used The aqueous solution elution of 90%ACN/1% formic acid), further ground with ethanol, obtain the chloro- N- of (4S) -8- (6- (((R) -2,2- bis- Methyl-1,3-dioxy heterocyclic pentane -4- bases) methoxyl group) pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (6.0g, 10.14mmol, 28.7% yield), It is white solid, LCMS (m/z):591.25[M+H]⁺。

Synthesizing the chloro- N- of (4S) -8-, (4- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) is phonetic Pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

NaH (0.283g, 5.89mmol) is added into the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4 in room temperature, 5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(1.0g, 2.94mmol) is at tetrahydrofuran (THF) In solution in (15mL), stir 1h and be slowly added to (S)-(4- ((2,2- dimethyl -1,3- dioxolane -4- bases) first Epoxide) pyrimidine -2-base) phenyl carbamates (2.033g, 5.89mmol), then stir 18h at 70 DEG C.By reactant mixture 28 DEG C are cooled to, is then distributed between water (20mL) and EtOAc (50mL).Organic layer is separated, then through anhydrous Na₂SO₄It is dry It is dry, filter and filtrate evaporation is obtained into crude product.Crude compound purifies (post through combiflash column chromatographys：C18, with 90% The aqueous solution elution of ACN/1% formic acid), further purify (silica gel through column chromatography：100-200 mesh, is washed with 90%EtOAc/ hexanes It is de-), obtain the chloro- N- of (4S) -8- (4- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -2- Base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (400mg, 0.659mmol, 22.38% yield), it is white solid LCMS (m/z), 591.50 (M+H)⁺。

Synthesize (4S) -8- chloro- N- (6- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

By the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza(15g, 44.2mmol) is dissolved in tetrahydrofuran (THF) (200mL), at 0 DEG C under nitrogen gas stirring Add triphosgene (10.48g, 35.3mmol), triethylamine (30.8mL, 221mmol).Reactant mixture is stirred at room temperature 30min.(S) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- amine is added thereto (14.85g, 66.2mmol) and stir 16h at 80 DEG C.So that reactant mixture reaches room temperature and gone out with 500ml water quenchings, use 3x800ml ethyl acetate is extracted.The organic layer of merging is through Na₂SO₄Dry and be concentrated under reduced pressure, obtain crude product.Crude product is through quick Column chromatography purifies (silica gel：100-200 mesh), obtain the chloro- N- of (4S) -8- (6- (((S) -2,2- dimethyl -1,3- dioxanes penta Alkane -4- bases) methoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza- 5 (2H)-formamides (9.3g, 15.66mmol, 35.5% yield), it is pale solid, LCMS (m/z):590.16[M+H]⁺。

Synthesizing the chloro- N- of (4S) -8-, (4- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) is phonetic Pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

At 0 DEG C to the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diazaIn the solution of (500mg, 1.472mmol) in THF (30mL) add triphosgene (218mg, 0.736mmol), then stir to room temperature, continue 1h.Then DIPEA (0.771mL, 4.42mmol) and (R) -4- are sequentially added ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -2- amine (663mg, 2.94mmol), in seal pipe Under the conditions of 75 DEG C keep 16h.Pass through TLC and LCMS monitoring reactions.Reactant mixture is poured into saturation NaHCO₃Solution In (50mL) and it is extracted with ethyl acetate (2x100mL).Organic layer is through anhydrous Na₂SO₄Dry and be concentrated under reduced pressure, obtain thick material. Thick material purifies (100-200 silica gel) through column chromatography, and the gradient mixture using 5% methanol/DCM is obtained as eluant, eluent The chloro- N- of (4S) -8- (4- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -2-base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (200mg, 0.335mmol, 22.77% yield), it is white solid LCMS (m/z):591.38[M+H]⁺。

Synthesize (4S) -8- chloro- N- (5- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Piperazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

In room temperature to the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diazaIn the solution of the stirring of (500mg, 1.472mmol) in tetrahydrofuran (THF) (20mL) Triphosgene (437mg, 1.472mmol) is added, triethylamine (1.231mL, 8.83mmol) is subsequently added into.Reactant mixture is stirred 45min, then adds (S) -5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- amine The solution of (497mg, 2.208mmol) in tetrahydrofuran (THF) (5mL).Reactant mixture is stirred into 12hr at 65 DEG C.TLC Indicate that initiation material exhausts and forms new spot.Water (25mL) is added into reactant mixture.Water layer is extracted with EtOAc (2 × 25mL).The organic layer of merging is through anhydrous Na₂SO₄It is dried, filtered and concentrated, obtains desired crude product.Purified on column chromatography Purifying, using 100-200 silica gel (eluant, eluent 35-50%EtOAc/ petroleum ethers), obtains desired pure product (4S) -8- chloro- N- (5- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- bases) -7- (3- (trifluoromethyl) Phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (500mg, 0.834mmol, 56.7% yield), it is pale solid.LCMS(m/z):591.16[M+H]⁺。

Synthesizing the chloro- N- of (4S) -8-, (2- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) is phonetic Pyridine -4- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

By the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza(30g, 88mmol), triphosgene (26.2g, 88mmol) and triethylamine (61.5mL, 442mmol) are four 15min is stirred at room temperature in solution in hydrogen furans (THF) (300mL) under a nitrogen.(S) -2- is added into the reactant mixture ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -4- amine (59.7g, 265mmol).By reaction mixing Thing stirs 16h at 70 DEG C, and monitors reaction process by TLC.Reactant mixture is cooled to room temperature, poured into water (200mL) And (3 × 200mL) is extracted with EtOAc.The organic layer of merging is washed with water (200mL), saline solution (200mL), through Na₂SO₄It is dry It is dry, filter and evaporate and obtain crude compound.Crude compound is purified through column chromatography, using neutral alumina and uses 20% EtOAc/ petroleum ethers are eluted, and obtain the pure chloro- N- of (4S) -8- (2- (((S) -2,2- dimethyl -1,3- dioxolane -4- Base) methoxyl group) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (23g, 38.8mmol, 44.0% yield), it is pale solid, LCMS (m/z): 591.4[M+H]⁺

Synthesizing the chloro- N- of (4S) -8-, (6- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) is phonetic Pyridine -4- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

TEA (0.821mL, 5.89mmol) is added into the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3 in room temperature, 4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(0.4g, 1.177mmol) is in tetrahydrofuran (THF) in (50mL) in the solution of stirring, triphosgene (0.349g, 1.177mmol) and stirring then are added in identical temperature 1h.Addition (S) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -4- amine (0.796g, 3.53mmol) and at 65 DEG C stir 15h.It is cooled to room temperature and is diluted with ethyl acetate (100mL) and water (100mL).Separation Organic layer is washed with water (50mL) and salt solution (50mL).Organic layer is through Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain roughization Compound.Purified and eluted with 50% ethyl acetate/hexane through column chromatography using aluminum oxide, obtain the chloro- N- (6- of (4S) -8- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine-4-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (0.25g, 0.414mmol, 35.2% yield), it is white solid, LCMS (m/z):591.1[M+H]⁺。

Synthesize (4S) -8- chloro- N- (4- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

Triethylamine (24.62mL, 177mmol) and triphosgene (8.73g, 29.4mmol) added into (4S) -8- in room temperature chloro- 7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza In the solution of the stirring of (10g, 29.4mmol) in THF.Reactant mixture is stirred into 45min and (R) -4- ((2,2- bis- are added Methyl-1,3-dioxy heterocyclic pentane -4- bases) methoxyl group) pyridine -2- amine (13.20g, 58.9mmol).By reactant mixture 65 DEG C stirring 16hr.Reactant mixture is cooled to room temperature, solvent is evaporated under reduced pressure completely, is then distributed in water (100mL) and EtOAc Between (500mL).Organic layer is separated, through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain crude product.Crude product is used Neutral alumina is purified through column chromatography, and is eluted with 30%EtOAc/ hexanes (gradient system), obtains desired product (8.50g), it is white solid.Product (8.50g) dilutes in ethanol (100mL), and with Silicycle palladium scavengers (4.25g) processing, then stirs 3hr at 65 DEG C.Reactant mixture is filtered by Celite pad, and by the hot second of Celite pad Alcohol (50ml) is washed, and the filtrate decompression of acquisition is concentrated, the chloro- N- of desired product (4S) -8- (4- (((R) -2,2- diformazans are obtained Base -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (8.0g, 13.52mmol, 45.9% yield), its For white solid, LCMS (m/z):590.07[M+H]⁺。

Synthesize (4S) -8- chloro- N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -4- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

At 0 DEG C to the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- bridges under nitrogen gas stirring Methylene pyridine simultaneously [2,3-b] [1,4] diaza(25g, 73.6mmol), triphosgene (13.10g, 44.2mmol) is in tetrahydrochysene DIPEA (64.3mL, 368mmol) is added in solution in furans (THF) (400mL).Then reactant mixture is stirred at 30 DEG C Mix 30min and add (R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- amine (18.15g, 81mmol), then stirs 16h by reactant mixture at 70 DEG C.Reaction is monitored through LCMS and TLC.Reaction is mixed Compound is poured into cold water (100mL) and is extracted with ethyl acetate (2x300mL).Organic layer is through anhydrous Na₂SO₄Dry and vacuum is dense Contracting.Crude compound obtains the chloro- N- of (4S) -8- through purification by flash chromatography (100-200 mesh, 90% ethyl acetate/petroleum ether) (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridin-4-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (18.5g, 30.9mmol, 41.9% yield), it is yellow solid.LCMS(m/z):590.12[M+H]⁺。

Synthesize (4S) -8- chloro- N- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -3- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

TEA (1.026mL, 7.36mmol) and triphosgene (437mg, 1.472mmol) are successively added into solution in room temperature The chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two AzepineIn the solution of (500mg, 1.472mmol) in tetrahydrofuran (THF) (20mL) and stirring 1h, add (R) -6- ((2, 2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -3- amine (330mg, 1.472mmol), then add at 80 DEG C Hot 15h.Reactant mixture is cooled to 28 DEG C, then distributed between water (25mL) and EtOAc (30mL × 3).Separation is organic Layer, then through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into thick material, then further purify it through column chromatography (using 100-200 silicagel columns, eluted with 50% ethyl acetate/hexane), obtains the chloro- N- of (4S) -8- (6- (((R) -2,2- diformazans Base -1,3- dioxolane -4- bases) methoxyl group) pyridin-3-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (350mg, 0.593mmol, 40.3% yield), It is pale solid, LCMS (m/z):590.16[M+H]⁺。

Synthesize (4S) -8- chloro- N- (5- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

DIPEA (571mg, 4.42mmol) and triphosgene (437mg, 1.472mmol) are added into (S) -5- in 25 DEG C of priorities ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- amine (660mg, 2.94mmol) is in tetrahydrofuran (THF) in the solution in (20mL), stir 1h and add the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetra- Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(500mg, 1.472mmol), then in 70 DEG C of heating 18hr.Reactant mixture is cooled to 28 DEG C, then distributed between water (20mL) and EtOAc (50mL).Organic layer is separated, so By anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into thick material (TLC eluant, eluents：100% ethyl acetate Rf:0.3；UV Activation).Crude compound purifies (C-18 through column chromatography:Eluted with the aqueous formic acid of 70% methanol/1%), obtain (4S) -8- Chloro- N- (5- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- bases) -7- (3- (fluoroforms Base) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (200mg, 0.339mmol, 23.03% yield), it is brownish thick thing, LCMS (m/z) 590.43 (M+H)⁺。

Synthesize (4S) -8- chloro- N- (5- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -3- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

At 0 DEG C by the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza(8g, 23.55mmol), TEA (16.41mL, 118mmol) is absorbed at tetrahydrofuran (THF) In (80mL), 10min is stirred at room temperature in gained yellow solution.Then 0 DEG C it is disposable add triphosgene (6.99g, 23.55mmol).Gained yellow suspension is stirred into 45min in room temperature.At 0 DEG C by (R) -5- ((2,2- dimethyl -1,3- dioxies Heterocycle pentane -4- bases) methoxyl group) THF (10mL) solution of pyridine -3- amine (7.92g, 35.3mmol) adds to above-mentioned yellow and is suspended Liquid, lasts 2min a period of time.Gained yellow suspension is heated to 70 DEG C, 24hr is kept.Reaction process is through TLC 5% MeOH/DCM is monitored, and TLC indicates to be formed initiation material after multiple spots, 24h and exhausted.Reactant mixture is cooled to room temperature, so After (20mL) is diluted with water, extracted (2 × 40mL), the organic layer of separation, washed with salt solution (20mL), through anhydrous with EtOAc Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain rough yellow solid.Thick material is purified through combiflash, uses silicagel column (12g, 5%MeOH/DCM).Fraction containing pure compound is merged and concentrated, the chloro- N- of desired compound (4S) -8- are obtained (5- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridin-3-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (11g, 18.53mmol, 79% yield), it is yellow solid, LCMS (m/z):590.06[M+H]⁺。

Synthesize (4S) -8- chloro- N- (6- (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethyoxyl) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first Acid amides

To the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaTriethylamine is added in the solution of (400mg, 1.177mmol) in tetrahydrofuran (THF) (30mL) (0.985mL, 7.06mmol) and triphosgene (349mg, 1.177mmol), is stirred at room temperature 30min under a nitrogen, to the reaction In mixture add 6- (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethyoxyl) pyridine -2- amine (701mg, 2.94mmol) and 16hr is stirred at 65 DEG C.TLC eluant, eluents：70% ethyl acetate/hexane R_f:0.3, UV activation.Reactant mixture is cooled to Room temperature, depressurizes evaporating completely solvent, then distributes between water (10mL) and EtOAc (2 × 50mL).Organic layer is separated, through nothing Water Na₂SO₄Dry, filter and evaporate filtrate, obtain thick material, it is brown solid.Thick material is diluted with DCM, Ran Houyong Neutral alumina absorbs and eluted with 25-30-%EtOAc/ petroleum ethers, collects fraction and concentrates, obtains the chloro- N- (6- of (4S) -8- (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethyoxyl) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(300mg, 0.478mmol, 40.6% are received -5 (2H)-formamides Rate), it is faint yellow solid, LCMS (m/z):604.14[M+H]⁺。

Synthesizing the chloro- N- of (4S) -8-, (5- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) is phonetic Pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

Triphosgene (437mg, 1.472mmol) is added into the chloro- 7- of (4S) -8- (3- (trifluoromethyl) at 28 DEG C under a nitrogen Phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(500mg, 1.472mmol) and In the solution of stirrings of the TEA (1.231mL, 8.83mmol) in tetrahydrofuran (THF) (50mL).By reactant mixture in room temperature 30min is stirred, and adds (S) -5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -2- amine (663mg,2.94mmol).By reactant mixture in 65 DEG C of stirrings.Reactant mixture is cooled to 28 DEG C；By reactant mixture point Fit between water (2mL) and EtOAc (2x25mL).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and steam filtrate Hair, obtains thick material.Thick material is purified through GRACE, C-18 reversed-phase columns, mobile phase A is used:0.1% formic acid/water；B:ACN, Product is with 50%ACN/0.1% formic acid/water elution.Evaporation solvent simultaneously uses saturation NaHCO₃Alkalization.The solid separated out is filtered, and is done It is dry, obtain the chloro- N- of (4S) -8- (5- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -2- Base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (300mg, 0.495mmol, 33.6% yield), it is pale solid, LCMS (m/z):591.19[M+H ]⁺。

Synthesizing the chloro- N- of (4S) -8-, (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) is phonetic Pyridine -5- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

Triphosgene (393mg, 1.325mmol) is added into the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2 at 28 DEG C, 3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(450mg, 1.325mmol) and triethylamine (1.108mL, 7.95mmol) is in triphosgene (393mg, 1.325mmol) in the solution of stirring.Reactant mixture is stirred into 2h And add (R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -5- amine (746mg, 3.31mmol).Reactant mixture is stirred into 10hr at 65 DEG C.Reactant mixture is cooled to room temperature, evaporating completely solvent is depressurized, Then distribute between water (40mL) and EtOAc (2 × 50mL).Organic layer is separated, through anhydrous Na₂SO₄Dry, filter and will filter Liquid evaporates, and obtains thick material, it is brown solid.Thick material is diluted with DCM, is then absorbed with neutral alumina and uses 40- 45%EtOAc/ petroleum ethers are eluted, and are collected fraction and are concentrated, obtain the chloro- N- of (4S) -8- (2- (((R) -2,2- dimethyl -1,3- Dioxolane -4- bases) methoxyl group) pyrimidine -5- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (450mg, 0.761mmol, 57.4% yield), it is ash White solid, LCMS (m/z):591.17[M+H]⁺。

Synthesize (4S) -8- chloro- N- (2- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -4- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

Triphosgene (21.84g, 73.6mmol) is added into the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2 at 25 DEG C, 3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(25.0g, 73.6mmol), and TEA In the solution of the stirring of (51.3mL, 368mmol) in tetrahydrofuran (THF) (200mL).Reactant mixture is stirred into 60min And add (S) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- amine (49.5g, 221mmol).Reactant mixture is stirred into 10hr at 72 DEG C.Reactant mixture is cooled to 25 DEG C, by the solid filtering of precipitation simultaneously Washed with ethyl acetate (100ml).Filtrate water (20ml) and saline solution (20ml) washing.Organic phase is separated, then through nothing Water Na₂SO₄Dry, filtering, and filtrate evaporation is obtained into thick material.By the thick material through neutral alumina purification by flash chromatography, use 30-40%EtOAc/ petroleum ethers are eluted, and obtain desired compound, it is solid.It is ground with 30% ether/pentane again Mill, obtains desired compound, it is pale solid.By the compound, (600mL) dilutes in ethanol, is used in combination Silicycle palladiums scavenger (12g) processing, then stirs 3hr at 50 DEG C.It is filtered by Celite pad, and by diatomite Pad is washed with hot ethanol (50ml), and the filtrate decompression of acquisition is concentrated, the chloro- N- of (4S) -8- (2- (((S) -2,2- diformazans are obtained Base -1,3- dioxolane -4- bases) methoxyl group) pyridin-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (22.0g, 37.1mmol, 50.4% yield), It is white solid, LCMS (m/z):590.07[M+H]⁺。

Synthesizing the chloro- N- of (4S) -8-, (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) is phonetic Pyridine -4- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

To the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaTriphosgene is added in the solution of (8.5g, 25.02mmol) in tetrahydrofuran (THF) (250mL) (7.42g, 25.02mmol), is subsequently added into triethylamine (20.92mL, 150mmol), and 20min is stirred at room temperature in gained suspension. By (R) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -4- amine (11.27g, 50.0mmol) Add in reactive material and gained suspension is heated to 70 DEG C, keep 16hr.By TLC monitorings reaction, (its display is not present Initiation material and in R_f:0.6 forms new spot).Thick material is through silica gel chromatography (100-200 silica gel, 0-3%MeOH/ DCM).Fraction containing pure compound is merged and concentrated, the chloro- N- of desired compound (4S) -8- (6- (((R) -2,2- are obtained Dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(8.5g, 13.19mmol, 52.7% are received -5 (2H)-formamides Rate), it is pale solid, LCMS (m/z):591.07(M+H)⁺。

Synthesize (4S) -8- chloro- N- (5- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

At 0 DEG C by the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza(500mg, 1.472mmol) tetrahydrofuran (THF) (15mL) and triphosgene (262mg, Solution in 0.883mmol) is stirred to room temperature, continues 30 minutes.Then successively add DIPEA (1.285mL, 7.36mmol) and (R) -5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- amine (578mg, 2.58mmol), so Afterwards 15hr is stirred at 75 DEG C 30 minutes.Pass through TLC and LCMS monitoring reactions.Reactant mixture is poured into saturation NaHCO₃Solution (50mL) and it is extracted with ethyl acetate (2x150mL).Organic layer is through anhydrous Na₂SO₄Dry and be concentrated under reduced pressure, obtain rough chemical combination Thing.Thick material purifies (100-200 silica gel) through column chromatography, and the gradient mixture using 80%EtOAc/ petroleum ethers is used as elution Agent, obtains the pure chloro- N- of compound (4S) -8- (5- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxies Base) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine- 5 (2H)-formamides (450mg, 0.691mmol, 47.0% yield), LCMS (m/z):590.37(M+H)⁺。

Synthesize (4S) -8- chloro- N- (6- (3- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) propoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first Acid amides

Triphosgene (437mg, 1.472mmol) is added into the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2 in room temperature, 3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(500mg, 1.472mmol) and TEA In the solution of the stirring of (1.231mL, 8.83mmol) in tetrahydrofuran (THF) (25mL).Reactant mixture is stirred into 4h, so 6- (3- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) propoxyl group) pyridine -2- amine (743mg, 2.94mmol) is added afterwards.Reaction is mixed Compound stirs 16h at 65 DEG C.Monitored and reacted by TLC.Reactant mixture is evaporated under reduced pressure, then in 500ml ethyl acetate Reconstruct.Organic layer is successively washed with water (200mL) and saline solution (100mL), uses anhydrous Na₂SO₄It is dried, filtered and concentrated, obtains To crude product.Crude product is determined by LCMS.Crude product is purified in next step, LCMS (m/z):618.10(M+H)⁺。

Synthesize (4S) -8- chloro- N- (6- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -3- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

DIPEA (571mg, 4.42mmol) and triphosgene (437mg, 1.472mmol) are added into (4S) -8- in 25 DEG C of priorities Chloro- 7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza In the solution of (500mg, 1.472mmol) in tetrahydrofuran (THF) (20mL), 1h is stirred, (S) -6- ((2,2- is then added Dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -3- amine (660mg, 2.94mmol), then in 70 DEG C of heating 18hr.Reactant mixture is cooled to 28 DEG C, then distributed between water (20mL) and EtOAc (50mL).Organic layer is separated, so By anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into thick material (TLC eluant, eluents：100% ethyl acetate R_f:0.3；UV Activation).Crude compound purifies (C-18 through column chromatography:Eluted with the aqueous formic acid of 70% methanol/1%), obtain (4S) -8- Chloro- N- (6- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridin-3-yl) -7- (3- (fluoroforms Base) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (350mg, 0.583mmol, 39.6% yield), it is brownish thick thing, LCMS (m/z):590.43(M+H)⁺。

Synthesize (4S) -8- chloro- N- (2- (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethyoxyl) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first Acid amides

TEA (0.821mL, 5.89mmol) and triphosgene (175mg, 0.589mmol) are successively added into the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza In the solution of the stirring of (400mg, 1.177mmol) in tetrahydrofuran (THF) (15mL), 1h is stirred at room temperature and 2- is added (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethyoxyl) pyrimidine -4- amine (310mg, 1.295mmol), then in 80 DEG C of heating 15h.Reactant mixture is cooled to 28 DEG C, then distributed between water (15mL) and EtOAc (30mL × 2).Separate organic layer, Then through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into thick material, be jelly.It is purified through column chromatography and (used 100-200 silica gel, post is eluted in 90% ethyl acetate/hexane), obtain the chloro- N- of (4S) -8- (2- (2- ((tetrahydrochysene -2H- pyrroles Mutter -2- bases) epoxide) ethyoxyl) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides (310mg, 0.505mmol, 42.9% yield), it is solid for canescence Body, LCMS (m/z):603.41(M+H)⁺。

Synthesizing the chloro- N- of (4S) -8-, (5- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) is phonetic Pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

By the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza(400mg, 1.177mmol), triphosgene (349mg, 1.177mmol) and triethylamine (0.821mL, 15min 5.89mmol) is stirred at room temperature under a nitrogen in the solution in tetrahydrofuran (THF) (20mL).Mixed to the reaction In thing add (R) -5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -2- amine (796mg, 3.53mmol).Reactant mixture is stirred into 16h at 70 DEG C, and reaction process is monitored by TLC.Reactant mixture is cooled to Room temperature, pours into water (10mL) and is extracted (3 × 20mL) with EtOAc.The organic layer of merging water (20mL), saline solution (20mL) Washing, through Na₂SO₄Dry, filter and evaporate and obtain crude compound.TLC eluant, eluents：100%EtOAc/ hexanes, R_f:0.1,UV Activation.Crude compound is purified through column chromatography, is eluted using neutral alumina and with 50%EtOAc/ petroleum ethers, is obtained pure The chloro- N- of (4S) -8- (5- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -2-base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (350mg, 0.590mmol, 50.1% yield), it is pale solid, LCMS (m/z):591.26[M+H]⁺。

Synthesizing the chloro- N- of (4S) -8-, (2- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) is phonetic Pyridine -5- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

TEA (0.985mL, 7.06mmol) and triphosgene (349mg, 1.177mmol) are added in room temperature under a nitrogen The chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two AzepineIn the solution of the stirring of (400mg, 1.177mmol) in tetrahydrofuran (THF) (40mL).Reactant mixture is existed 30min is stirred at room temperature.Add (S) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -5- amine (530mg, 2.355mmol) and reactant mixture is stirred into 15h at 65 DEG C.Reactant mixture is cooled to room temperature, decompression is complete Evaporation solvent, is then distributed between water (30mL) and EtOAc (100mL).Organic layer is separated, through anhydrous Na₂SO₄Dry, filtering And evaporate filtrate, obtain crude product.Purified on column chromatography is purified, and using neutral alumina and uses 30%EtOAc/ hexanes (gradient system) is eluted, and obtains desired product (the chloro- N- of (4S) -8- (2- (((S) -2,2- dimethyl -1,3- dioxanes penta Alkane -4- bases) methoxyl group) pyrimidine -5- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza- 5 (2H)-formamides (300mg, 0.495mmol, 42.1% yield), it is faint yellow solid (TLC Eluant, eluent：70%EtOAc/ hexanes:R_f0.5；UV activation), LCMS (m/z):591.19[M+H]⁺。

Synthesize (4S) -8- chloro- N- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

At 0 DEG C to the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diazaIn the solution of (10g, 29.4mmol) in THF (300mL) add triphosgene (4.37g, 14.72mmol), then stir to room temperature, continue 1h.Then successively add TEA (20.51mL, 147mmol) and (R) -6- ((2, 2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- amine (13.20g, 58.9mmol), in seal pipe condition Under 75 DEG C keep 16h.Pass through TLC and LCMS monitoring reactions.Reactant mixture is poured into saturation NaHCO₃Solution (300mL) is simultaneously It is extracted with ethyl acetate (1000mL).Organic layer is through anhydrous Na₂SO₄Dry and be concentrated under reduced pressure, obtain thick material.Thick material is through post Chromatogram purification (100-200 silica gel), the gradient mixture using 80%EtOAc/ petroleum ethers obtains (4S) -8- as eluant, eluent Chloro- N- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- bases) -7- (3- (fluoroforms Base) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (8.2g, 13.81mmol, 46.9% yield).Then by the material through Pd scavenger resin process purifications.At 50 DEG C to the chloro- N- of (4S) -8- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (8.2g) are in ethanol In stirring in (250ml) solution in add Pd scavenger resins (4.5g) and stir 5h at 70 DEG C.40 DEG C are subsequently cooled to, Reactant mixture is filtered and is concentrated under reduced pressure, (the chloro- N- of (4S) -8- (6- (((R) -2,2- dimethyl -1,3- dioxanes are obtained Pentane -4- bases) methoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (8g), it is white solid, LCMS (m/z) 590.12 [M+H]⁺。

Synthesize (4S) -8- chloro- N- (4- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Triethylamine (1.312ml, 9.42mmol) and triphosgene (466mg, 1.569mmol) are added in room temperature under a nitrogen The chloro- 7- of (4S) -8- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneousIn the solution of the stirring of (450mg, 1.569mmol) in tetrahydrofuran (THF) (30mL).By reactant mixture in room Temperature stirring 30min.Add (R) -4- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- amine (704mg, 3.14mmol) and reactant mixture is stirred into 16h at 65 DEG C.Reactant mixture is cooled to room temperature；Decompression is steamed completely Solvent is sent out, is then distributed between water (30mL) and EtOAc (100mL).Organic layer is separated, through anhydrous Na₂SO₄Dry, filtering is simultaneously Filtrate is evaporated, thick material is obtained, it is brown solid.Thick material is purified through column chromatography, using neutral alumina and uses 50% EtOAc/ Hex, obtains the chloro- N- of desired product (4S) -8- (4- (((R) -2,2- dimethyl -1,3- dioxanes penta Alkane -4- bases) methoxyl group) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza- 5 (2H)-formamides (450mg, 0.383mmol, 24.40% yield), it is faint yellow solid, LCMS(m/z):536.9[M+H]⁺。

Synthesize (4S) -8- chloro- N- (5- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Piperazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to the chloro- 7- of (4S) -8- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diazaAdd in the solution of the stirring of (500mg, 1.744mmol) in tetrahydrofuran (THF) (10mL) Enter triphosgene (517mg, 1.744mmol), be subsequently added into triethylamine (1.458mL, 10.46mmol).Reactant mixture is stirred 45min, then adds (S) -5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- amine The solution of (785mg, 3.49mmol) in tetrahydrofuran (THF) (5mL).Reactant mixture is stirred into 16hr at 65 DEG C.Pass through TLC monitors reaction process.TLC indicates that initiation material is depleted.Reactive material is cooled down to room temperature, (50mL) is diluted with water and second is used Acetoacetic ester extracts (50mL × 2).Merge organic layer and through Na₂SO₄It is dried, filtered and concentrated, obtains thick material, it is viscous for brown Thick compound.Crude product is purified in combiflash silicagel columns (40g) and eluted with Hex/EtOAc.Collect fraction:50% EtOAc/ petroleum ethers, eluted product.Enriched product fraction, obtains the chloro- N- of (4S) -8- (5- (((S) -2,2- dimethyl -1,3- bis- Tetrahydrofuran -4- bases) methoxyl group) pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (250mg, 0.447mmol, 25.6% yield), it is light yellow Solid.

LCMS(m/z):538.14[M+H]⁺。

Synthesize (4S) -8- chloro- N- (5- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Piperazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to the chloro- 7- of (4S) -8- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diazaAdd in the solution of the stirring of (500mg, 1.744mmol) in tetrahydrofuran (THF) (5mL) Enter triphosgene (517mg, 1.744mmol), 30min is stirred at room temperature in reactant mixture, then by (R) -5- ((2,2- diformazans Base -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- amine (589mg, 2.62mmol) adds to reactant mixture, and will be anti- Mixture is answered to stir 18hr at 70 DEG C.Reaction process is monitored by TLC.Reactant mixture is diluted with water (30mL) and extracted with EtOAc Take (3 × 30mL), organic layer merges and uses aqueous salt solu-tion (30mL), and organic layer is through anhydrous Na₂SO₄Dry, filter and depressurize Concentration, is obtained crude compound, thick material is purified through column chromatography, is washed using 100-200 mesh silica gel and with 40%EtOAc/ hexanes De- compound, obtains the chloro- N- of (4S) -8- (5- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Piperazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides (450mg, 0.807mmol, 46.3% yield), it is faint yellow solid.LCMS(m/z):538.47(M+ H)⁺。

Synthesize (4S) -8- chloro- N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -4- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- 7- of (4S) -8- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- bridges under nitrogen gas stirring Methylene pyridine simultaneously [2,3-b] [1,4] diaza(350mg, 1.221mmol), triphosgene (217mg, 0.732mmol) exists Triphosgene (217mg, 0.732mmol) is added in solution in tetrahydrofuran (THF) (15mL).Then by reactant mixture 30 DEG C stirring 30min simultaneously adds (R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- amine (411mg, 1.831mmol), then stirs 15h 30min by reactant mixture at 70 DEG C.Reactant mixture is monitored through LCMS.Will Reactant mixture is poured into cold water (20mL) and is extracted with ethyl acetate (2x20mL).Organic layer is through anhydrous Na₂SO₄Dry and true Sky concentration, obtains crude product.Crude compound obtains the chloro- N- (2- of (4S) -8- through purification by flash chromatography (100-200 mesh) (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridin-4-yl) -7- (2- picoline -4- bases) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (450mg, 0.778mmol, 63.7% yield), it is yellow solid, LCMS (m/z):536.9[M+H]⁺。

Synthesize (4S) -8- chloro- N- (4- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- 7- of (4S) -8- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- bridges under nitrogen gas stirring Methylene pyridine simultaneously [2,3-b] [1,4] diaza(250mg, 0.872mmol), triphosgene (155mg, 0.523mmol) exists DIPEA (0.761mL, 4.36mmol) is added in solution in tetrahydrofuran (THF) (20mL).Then by reactant mixture 30 DEG C stirring 30min simultaneously adds (S) -4- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- amine (293mg, 1.308mmol), then stirs 15h 30min by reactant mixture at 70 DEG C.Reaction is monitored through LCMS and TLC. Reactant mixture is poured into cold water (20mL) and is extracted with ethyl acetate (2x50mL).Organic layer is through anhydrous Na₂SO₄Dry and It is concentrated in vacuo.Crude compound obtains (4S) -8- through purification by flash chromatography (100-200 mesh, 90% ethyl acetate/petroleum ether) Chloro- N- (4- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- bases) -7- (2- methyl pyrroles Pyridine -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (230mg, 0.417mmol, 47.9% yield), LCMS (m/z):537.05[M+H]⁺。

Synthesize (4S) -8- chloro- N- (6- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the chloro- 7- of (4S) -8- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaTriphosgene is added in the solution of (400mg, 1.395mmol) in tetrahydrofuran (THF) (30mL) (400mg, 1.348mmol), 20min is stirred at room temperature in resulting solution.By (S) -6- ((2,2- dimethyl -1,3- dioxanes penta Alkane -4- bases) methoxyl group) solution of the pyridine -2- amine (626mg, 2.79mmol) in THF (10mL) adds to reactive material and by institute Obtain suspension and be heated to 60 DEG C, keep 16hr.(monitored after the completion of reaction through TLC, it was observed that polarity and nonpolar two spots Point), water (20ml) is added into reactive material and is extracted with ethyl acetate (2X20ml).Organic layer is through Na₂SO₄Dry filter simultaneously subtracts Pressure concentration, obtains dark brown liquid.Thick material is purified through combiflash, uses silicagel column (12g, 80%EtOAc/ oil Ether).Fraction containing pure compound is merged and concentrated, the chloro- N- of desired compound (4S) -8- (6- (((S) -2,2- are obtained Dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(330mg, 0.596mmol, 42.7% are received -5 (2H)-formamides Rate), it is Light brown solid.LCMS(m/z):537.2(M+H)⁺。

Synthesizing the chloro- N- of (4S) -8-, (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) is phonetic Pyridine -4- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Triethylamine (1.458mL, 10.46mmol) and triphosgene (517mg, 1.744mmol) are added into (4S) -8- in room temperature Chloro- 7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza In the solution of the stirring of (500mg, 1.744mmol) in tetrahydrofuran (THF) (25mL), 30min is stirred.By (R) -6- ((2, 2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -4- amine (589mg, 2.62mmol) add to above-mentioned reaction mix Compound, 16h is stirred at 70 DEG C.Reactant mixture is reached room temperature, removed by rotary evaporation from reactant mixture organic molten Agent, by residue distribution between water (40mL) and EtOAc (80mL).Separate organic layer and through anhydrous Na₂SO₄Dry, filtering is simultaneously Filtrate evaporation is obtained into crude compound.Crude compound is purified through column chromatography, uses silica gel (100-200 mesh), 40% acetic acid Ethyl ester/hexane obtains the chloro- N- of (4S) -8- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- as eluant, eluent Base) methoxyl group) pyrimidine-4-yl) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (225mg, 0.383mmol, 21.94% yield), it is faint yellow solid.(TLC Eluant, eluent：5%MeOH/DCM R_f:0.4；UV activation) .LCMS (m/z):538.28[M+H]⁺。

Synthesizing the chloro- N- of (4S) -8-, (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) is phonetic Pyridine -4- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the chloro- 7- of (4S) -8- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaTriethylamine is added in the solution of (500mg, 1.744mmol) in tetrahydrofuran (THF) (30mL) (1.458mL, 10.46mmol) and triphosgene (517mg, 1.744mmol).30min is stirred at room temperature in reactant mixture.To this (R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -4- amine is added in reactant mixture (1178mg, 5.23mmol) and pass through TLC monitoring reactions in 65 DEG C of stirring.Removal of solvent under reduced pressure, is diluted with water (20mL) and is used in combination Ethyl acetate extracts (2 × 50mL).The organic layer of merging water (30mL), saturated brine solution (10mL) washing, through anhydrous sulphur Sour sodium is dried, filtered and concentrated.Crude compound is dissolved in DCM (15mL).Neutral alumina is added into crude compound simultaneously Purified through column chromatography.Product is eluted with 30-35% ethyl acetate/hexanes.The fraction of collection is evaporated under reduced pressure, obtains pure The chloro- N- of (4S) -8- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine-4-yl) -7- (2- Picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (200mg, 0.370mmol, 21.23% yield), it is pale solid, LCMS (m/z):536.25(M+H)⁺。

Synthesize (4S) -8- chloro- N- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Piperazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

TEA (0.729mL, 5.23mmol) and triphosgene (517mg, 1.744mmol) are added into (4S) -8- in 25 DEG C of priorities Chloro- 7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza In the solution of (500mg, 1.744mmol) in tetrahydrofuran (THF) (20mL), stir 1h and add (R) -6- ((2,2- diformazans Base -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- amine (785mg, 3.49mmol), then heats 15h at 70 DEG C. Reactant mixture is cooled to 28 DEG C, then distributed between water (50mL) and EtOAc (100mL).Separate organic layer, Ran Houjing Anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into thick material, be jelly (TLC eluant, eluents：Net ethyl acetate R_f:0.4； UV activation).Purified on column chromatography purifies (silica gel：100-200 mesh, uses 90%EtOAc/ Hex), obtain (4S) -8- Chloro- N- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- bases) -7- (2- methyl pyrroles Pyridine -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (250mg, 0.428mmol, 24.53% yield), it is pale solid, LCMS (m/z):538.35(M+H)⁺。

Synthesize (4S) -8- chloro- N- (6- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Piperazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the chloro- 7- of (4S) -8- (2- picoline -4- the bases) -2,3,4,5- tetrahydrochysenes being stirred at room temperature in a nitrogen atmosphere - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(500mg, 1.744mmol) is at tetrahydrofuran (THF) (50mL) In solution in add triethylamine (1.458mL, 10.46mmol) and triphosgene (517mg, 1.744mmol).By reactant mixture 30min is stirred at room temperature, be subsequently added (S) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine - 2- amine (785mg, 3.49mmol).Then reactant mixture is stirred into 16h at 65 DEG C.Monitored and reacted by TLC.By reaction mixing Thing is evaporated under reduced pressure, and is then reconstructed in 200ml ethyl acetate.Organic layer is successively washed with water (100mL) and saline solution (50mL) Wash and use Na₂SO₄Drying and dehydrating, filters and concentrates, obtain crude product.Crude product carries out silica gel (100-200 mesh), and uses 100% Ethyl acetate is eluted, and obtains the chloro- N- of compound (4S) -8- (6- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) Methoxyl group) pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] Diaza- 5 (2H)-formamides (230mg, 0.396mmol, 22.72% yield), it is white solid, LCMS (m/z): 537.9(M+H)⁺。

Synthesize (4S) -8- chloro- N- (2- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -4- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Triphosgene (414mg, 1.395mmol) is added into the chloro- 7- of (4S) -8- (2- picoline -4- bases) -2,3 at 28 DEG C, 4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(400.0mg, 1.395mmol), and TEA In the solution of the stirring of (0.972mL, 6.97mmol) in tetrahydrofuran (THF) (20.0mL).Reactant mixture is stirred 30min and add (S) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- amine (938mg, 4.18mmol).Reactant mixture is stirred into 10hr at 72 DEG C.Reactant mixture is cooled to 25 DEG C, the solid of precipitation is filtered And (40mL) is washed with ethyl acetate.Filtrate water (10ml) and saline solution (10mL) washing.Organic phase is separated, then through nothing Water Na₂SO₄Dry, filtering, and filtrate evaporation is obtained into thick material.By the thick material through neutral alumina purification by flash chromatography, use 20-30%EtOAc/ petroleum ethers elute, obtain the chloro- N- of (4S) -8- (2- (((S) -2,2- dimethyl -1,3- dioxolane - 4- yls) methoxyl group) pyridin-4-yl) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (380.0mg, 0.677mmol, 48.5% yield), it is white solid, LCMS (m/z):537.34(M+H)⁺。

Synthesize (4S) -8- chloro- N- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Triphosgene (414mg, 1.395mmol) is added into the chloro- 7- of (4S) -8- (2- picoline -4- bases) -2,3 in room temperature, 4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(400mg, 1.395mmol) is in tetrahydrofuran (THF) in the solution of the stirring in (10mL).45min is stirred at room temperature in reactant mixture, triethylamine is then sequentially added (1.167mL, 8.37mmol) and (R) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- amine (626mg,2.79mmol).Reactant mixture is stirred into 6hr at 65 DEG C.Monitored and reacted by TLC.TLC shows a polarity spot Put and initiation material is depleted.Stop reaction.Reactant mixture is concentrated under reduced pressure into dry.Residue is absorbed at DCM (100mL) In, by organic layer priority water and aqueous salt solu-tion.Organic layer is through Na₂SO₄It is dried, filtered and concentrated, obtains crude product.Slightly Product is through silica gel chromatography (100-200 mesh), and by post 30%EtOAc/ Hex.Collect pure fraction and steaming Hair, obtains required product (the chloro- N- of (4S) -8- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (300mg, 0.531mmol, 38.0% yield), it is white solid, LCMS (m/z):537.2[M+ H]⁺。

Synthesizing the chloro- N- of (4S) -8-, (6- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) is phonetic Pyridine -4- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

TEA (0.972mL, 6.97mmol) is added into the chloro- 7- of (4S) -8- (2- picoline -4- bases) -2,3,4 in room temperature, 5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(0.4g, 1.395mmol) is at tetrahydrofuran (THF) In the solution of stirring in (50mL), then triphosgene (0.414g, 1.395mmol) and stirring 1h are added in identical temperature. Addition (S) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -4- amine (0.943g, 4.18mmol), then 15h is stirred at 65 DEG C.Room temperature is cooled to, is then diluted with ethyl acetate (50mL) and water (50mL).Point From organic layer washed with water (50mL) and salt solution (50mL).Organic layer is through Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain thick Produced compounds.Crude compound is purified by column chromatography, is eluted, is obtained using aluminum oxide and in 50% ethyl acetate/hexane The chloro- N- of (4S) -8- (6- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine-4-yl) -7- (2- Picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (0.350g, 0.644mmol, 46.2% yield), it is white solid, LCMS (m/z):538.28[M+H]⁺。

Synthesizing the chloro- N- of (4S) -8-, (2- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) is phonetic Pyridine -4- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

By the chloro- 7- of (4S) -8- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza(300mg, 1.046mmol), triphosgene (310mg, 1.046mmol) and triethylamine (0.875mL, 6.28mmol) 15min is stirred at room temperature in the solution in tetrahydrofuran (THF) (5mL) under a nitrogen.Into the reactant mixture Addition (S) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -4- amine (707mg, 3.14mmol).Reactant mixture is stirred into 16hr at 70 DEG C, and reaction process is monitored by TLC.Reactant mixture is cooled to Room temperature, pours into water (10mL) and is extracted (3 × 20mL) with EtOAc.Then by the organic layer of merging with water (10mL), salt is water-soluble Liquid (10mL) is washed, through Na₂SO₄Dry and evaporate and obtain crude compound.Crude compound is purified through column chromatography, uses neutrality Aluminum oxide and eluted with 100%EtOAc, obtain the chloro- N- of (4S) -8- (2- (((S) -2,2- dimethyl -1,3- dioxanes penta Alkane -4- bases) methoxyl group) pyrimidine-4-yl) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza- 5 (2H)-formamides (250mg, 0.379mmol, 36.2% yield), LCMS (m/z):537.9[M+ H]⁺。

Synthesize (4S)-N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- Base) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide

In room temperature to solid (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- bridges under nitrogen gas stirring Methylene pyridine simultaneously [2,3-b] [1,4] diazaThe solution of (400mg, 1.585mmol) in THF (30mL) adds solid Triphosgene (282mg, 0.951mmol), is stirred at room temperature 30 minutes under a nitrogen.Then thereto add DIPEA (1.384mL, 7.93mmol) and (R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- amine (533mg, 2.378mmol), 16h is kept at 75 DEG C under the conditions of seal pipe.Pass through TLC and LCMS monitoring reactions.Reactant mixture is dense Contracting, DCM (200mL) is absorbed in by residue.By solution water and salt water washing, through Na₂SO₄It is dried, filtered and concentrated, obtains Crude compound.Crude product is added in neutral alumina, and is eluted with 50%EtOAc/ petroleum ethers.Collect fraction and concentrate, again The impure compound of identical is obtained, is purified with preparation HPLC, pure compound (4S)-N- (2- (((R) -2,2- bis- are obtained Methyl-1,3-dioxy heterocyclic pentane -4- bases) methoxyl group) pyridin-4-yl) -7- (2- picoline -4- bases) -3,4- dihydro -1, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(150mg, 0.298mmol, 18.83% are received -5 (2H)-formamides Rate), LCMS (m/z):503.39[M+H]⁺。

Synthesize (4S)-N- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- Base) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide

In room temperature to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- bridge methylenes under nitrogen gas stirring Yl pyridines simultaneously [2,3-b] [1,4] diazaIn the solution of (300mg, 1.189mmol) in tetrahydrofuran (THF) (5mL) Add triphosgene (353mg, 1.189mmol) and TEA (0.166mL, 1.189mmol) and stir 2h, (R) -6- is added thereto ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- amine (268mg, 1.189mmol) adds and will Reactant mixture stirs 16hr at 60 DEG C.Reactant mixture is quenched with frozen water and uses 2x15ml ethyl acetate to extract, and merging has Machine layer 10ml water and 5ml aqueous salt solu-tions, organic layer is through Na₂SO₄Dry and be concentrated under reduced pressure, obtain crude compound, LCMS (m/z):504.40[M+H]⁺。

Synthesize (4S)-N- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- Base) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide

In room temperature to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- bridge methylenes under nitrogen gas stirring Yl pyridines simultaneously [2,3-b] [1,4] diazaIn the solution of (500mg, 1.982mmol) in tetrahydrofuran (THF) (15mL) Triphosgene (588mg, 1.982mmol) and TEA (1.657mL, 11.89mmol) stirring 2h are added, (R) -6- is added thereto ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- amine (889mg, 3.96mmol) is simultaneously mixed by reaction Compound stirs 16hr at 60 DEG C.Monitored and reacted by TLC.Reactant mixture is quenched with frozen water and uses 2x15ml ethyl acetate to extract Take, organic layer the 15ml water and 15ml aqueous salt solu-tions of merging, organic layer is through Na₂SO₄Dry and be concentrated under reduced pressure, obtain thick Produced compounds.Crude product is added into 100-200 silicagel columns and eluted with 2% DCM/MeOH, pure compound (4S)-N- is obtained (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- bases) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (350mg, 0.612mmol, 30.9% yield), it is faint yellow solid, LCMS (m/z):503.39[M+H]⁺。

Synthesize (4S)-N- (4- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- Base) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide

In room temperature to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- bridge methylenes under nitrogen gas stirring Yl pyridines simultaneously [2,3-b] [1,4] diazaIn the solution of (300mg, 1.189mmol) in tetrahydrofuran (THF) (10mL) Add triphosgene (353mg, 1.189mmol) and TEA (0.994mL, 7.13mmol) and stir 2h, (R) -4- is added thereto ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- amine (533mg, 2.378mmol) simultaneously will reaction Mixture stirs 16hr at 60 DEG C.Monitored and reacted by TLC.Reactant mixture is quenched with frozen water and uses 2x15ml ethyl acetate to extract Take, organic layer the 15ml water and 15ml aqueous salt solu-tions of merging, organic layer is through Na₂SO₄Dry and be concentrated under reduced pressure, obtain thick Produced compounds.Crude product is added into 100-200 silicagel columns and eluted with 2%DCM/MeOH, pure compound (4S)-N- is obtained (4- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- bases) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (200mg, 0.277mmol, 23.27% yield), it is Light brown solid, LCMS (m/z):503.45[M+H]⁺。

Synthesize (4S)-N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -4- Base) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide

Room temperature successively by triethylamine (1.381mL, 9.91mmol) and triphosgene (294mg, 0.991mmol) add to (R)- 2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -4- amine (580mg, 2.58mmol) is in tetrahydrochysene furan In the solution for the stirring muttered in (THF) (100mL) and stirring 5h.Then room temperature by (4S) -7- (2- picoline -4- bases) - 2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(500mg, 1.982mmol) adds to reaction Mixture, then stirs 15h at 80 DEG C.Reactant mixture is cooled to room temperature, (50mL) is diluted with water, is extracted with ethyl acetate (2 × 75mL) and with salt water washing (100mL).Organic layer is separated, through Na₂SO₄It is dried, filtered and concentrated, obtains rough chemical combination Thing, LCMS:(m/z):504.46[M+H]⁺。

Synthesize (4S)-N- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -4- Base) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide

Successively triphosgene (353mg, 1.189mmol) and triethylamine (1.657mL, 11.89mmol) are added in room temperature (R) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -4- amine (893mg, 3.96mmol) is four In the solution of stirring in hydrogen furans (THF) (20mL) and stirring 1h.Then by (4S) -7- (2- picoline -4- bases) -2,3, 4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(500mg, 1.982mmol) adds to reaction mixing Thing, then stirs 15h at 80 DEG C.Reactant mixture is cooled to room temperature, (50mL) is diluted with water, is extracted with ethyl acetate (2X70mL), with aqueous salt solu-tion (20mL).Organic layer is separated, through Na₂SO₄It is dried, filtered and concentrated, obtains rough chemical combination Thing, LCMS (m/z):504.28[M+H]⁺。

Synthesize (4R) -7- chloro- N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine- 5 (2H)-formamides

To suspension under a nitrogen in 20 DEG C of nicotinic acid (2.8g, 22.74mmol) stirred in tetrahydrofuran (100mL) Middle addition DPPA (9.39g, 34.1mmol), DIPEA (11.92mL, 68.2mmol) solution, are then stirred again in identical temperature 1h is mixed, chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of solid (4R) -7- are added (3.56g,18.20mmol).Reactant mixture is stirred into 16hr at 80 DEG C.Pass through TLC and LCMS monitoring reactions.By reaction mixing Thing is directly evaporated under reduced pressure, and is diluted, is subsequently washed with water with ethyl acetate (200mL).Organic layer is through anhydrous Na₂SO₄Dry, decompression Thick material is concentrated to give, (100-200 silica gel) is purified through column chromatography, the gradient mixture using 1% methanol/EtOAc is as washing De- agent, obtains the chloro- N- of (4R) -7- (pyridin-3-yl) -3,4- dihydros-Isosorbide-5-Nitrae-endo-methylene group pyrido [2,3-b] [Isosorbide-5-Nitrae] phenodiazine It is miscellaneous- 5 (2H)-formamides (4g, 12.13mmol, 53.3% yield), it is pale solid, LCMS (m/z):316.19[M+ H]⁺。

Synthesize (4R) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza

To chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4R) -7-(2g, 10.22mmol), (2- picoline -4- bases) boric acid (2.100g, 15.33mmol) and K₃PO₄(6.51g, 30.7mmol) 1, X-phos (1.949g, 4.09mmol), Pd are added in 4- dioxanes (40mL) and water (10mL) in the solution of degassing₂(dba)₃ (1.872g,2.044mmol).Reactant mixture is stirred into 3hr at 110 DEG C.Monitored and reacted by TLC.Reactant mixture is fallen Enter cold water (50mL) and be extracted with ethyl acetate (2x100mL).Organic layer is through anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure.Slightly Product adds to silicagel column and eluted with 2%DCM/MeOH.The fraction of collection is evaporated, (4R) -7- (2- picolines -4- are obtained Base) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(1.2g, 4.71mmol, 46.1% Yield), it is pale solid, LCMS (m/z):253.0[M+H]⁺。

Synthesize (4R)-N- (4- bromopyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to solid (4R) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- bridges under nitrogen gas stirring Methylene pyridine simultaneously [2,3-b] [1,4] diaza(1.2g, 4.76mmol) adds solid in tetrahydrofuran (THF) (80mL) Body triphosgene (0.847g, 2.85mmol), is stirred at room temperature 30 minutes under a nitrogen.Thereto add DIPEA (4.15mL, 23.78mmol) with 4- bromopyridine -2- amine (1.234g, 7.13mmol).Reactant mixture is stirred into 16hr at 75 DEG C.Pass through TLC Monitor and react with LCMS.(60ml) is diluted with water in reactant mixture and is extracted with ethyl acetate (3X100ml).Separate organic layer And through anhydrous Na₂SO₄Dry and be concentrated under reduced pressure, obtain crude product.By thick material by purification by flash chromatography, silica gel (100- is used 200 mesh), using 2.5% methanol-DCM as eluant, eluent, obtain pure compound (4R)-N- (4- bromopyridine -2- bases) -7- (2- Picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (1g, 2.194mmol, 46.1% yield), it is faint yellow solid LCMS (m/z) 453.26 [M+H]⁺。

To the bromo- 7- of (4S) -8- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza(1g, 3.02mmol), methyl-boric acid (0.361g, 6.04mmol) and K₃PO₄(1.923g, Solid Pd 9.06mmol) is added in the solution of the degassing in 1,4- dioxanes (20mL) and water (5mL)₂(dba)₃(0.276g, 0.302mmol) with x-phos (0.288g, 0.604mmol).Reactant mixture is stirred into 15.5hr at 100 DEG C.By reaction mixing Thing is diluted with water (50mL) and is extracted with ethyl acetate (3 × 50mL).Organic layer is through anhydrous Na₂SO₄Dry, be concentrated under reduced pressure to give Half pure compound.Crude product is purified through flash chromatography (100-200 mesh) and eluted with 3%DCM/EtOAc, obtains (4S) -8- Methyl -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza (500mg, 1.802mmol, 59.7% yield), it is yellow solid, LCMS (m/z):267.1[M+H]⁺。

Synthesize (4S) -8- bromo- 7- (2- picoline -4- bases)-N- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the bromo- 7- of (4S) -8- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysenes -1,4- under being stirred under a nitrogen at 0 DEG C Endo-methylene group pyrido [2,3-b] [1,4] diaza(600mg, 1.812mmol) is in tetrahydrofuran (THF) (20mL) NaH (65.2mg, 2.72mmol) is added in solution.Then reactant mixture is stirred into 1h at 30 DEG C, adds 3- (pyridine -2- Base) -2H- pyridos [1,2-a] [1,3,5] triazine -2,4 (3H)-diketone (522mg, 2.174mmol), then reaction is mixed Thing stirs 15h at 70 DEG C.Monitored through LCMS.Reactant mixture is poured into cold water (20mL) and is extracted with ethyl acetate (0mL). Organic layer is through anhydrous Na₂SO₄Dry and be concentrated in vacuo.Crude compound is through purification by flash chromatography (100-200 mesh, 2%MeOH/ DCM is used as eluant, eluent), obtain the bromo- 7- of desired product (4S) -8- (2- picoline -4- bases)-N- (pyridine -2- bases) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (320mg, 0.579mmol, 31.9% yield), it is white solid, LCMS (m/z):450.8[M+H]⁺。

Synthesize (4S)-N- (6- (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethyoxyl) pyrazine -2- bases) -7- (3- (three Methyl fluoride) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

TEA (1.141mL, 8.19mmol) and triphosgene (486mg, 1.638mmol) are successively added into (4S) -7- in room temperature (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza In the solution of (500mg, 1.638mmol) in tetrahydrofuran (THF) (20mL) and stirring 1h, then add 6- (2- ((tetrahydrochysene- 2H- pyrans -2- bases) epoxide) ethyoxyl) and pyrazine -2- amine (431mg, 1.802mmol) and 80 DEG C heat 15h.By reaction mixing Thing is cooled to 28 DEG C, then distributes water (25mL) and EtOAc (30mL × 2) between.Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into (4S)-N- (6- (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethyoxyl) pyrroles Piperazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides (310mg, 0.498mmol, 30.4% yield), it is pale solid, LCMS:571.35(M+H)⁺。

Synthesize (4S)-N- (2- (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethyoxyl) pyrimidine-4-yl) -7- (3- (three Methyl fluoride) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

TEA (1.141mL, 8.19mmol) and triphosgene (486mg, 1.638mmol) are successively added into (4S) -7- in room temperature (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza In the solution of (500mg, 1.638mmol) in tetrahydrofuran (THF) (25mL), then stir 1h and add 2- (2- ((tetrahydrochysene- 2H- pyrans -2- bases) epoxide) ethyoxyl) pyrimidine -4- amine (431mg, 1.802mmol), and heat 15h at 80 DEG C.Reaction is mixed Compound is cooled to 28 DEG C, then distributes between water (25mL) and EtOAc (25mL × 2).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained (4S)-N- (2- (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethyoxyl) is phonetic Pyridine -4- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides (300mg, 0.510mmol, 31.1% yield), it is pale solid, LCMS (m/z):571.11(M+ H)⁺。

Synthesize (4S)-N- (6- (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethyoxyl) pyridine -2- bases) -7- (3- (three Methyl fluoride) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diazaTriethylamine is added in the solution of (350mg, 1.146mmol) in tetrahydrofuran (THF) (20mL) (0.959mL, 6.88mmol) and triphosgene (340mg, 1.146mmol), is stirred at room temperature 1h under a nitrogen, is mixed to the reaction 6- (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethyoxyl) pyridine -2- amine (820mg, 3.44mmol) is added in thing and 65 DEG C stirring 16h.TLC eluant, eluents：70% ethyl acetate/hexane R_f:0.3, UV activation.Reactant mixture is cooled to room temperature； Evaporating completely solvent is depressurized, is then distributed between water (10mL) and EtOAc (2 × 50mL).Organic layer is separated, through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain thick material, it is brown solid.Thick material is diluted with DCM, in then using Property aluminum oxide absorb and with 30-35-%EtOAc/ petroleum ethers elute, collection fraction simultaneously concentrate, obtain (4S)-N- (6- (2- ((four Hydrogen -2H- pyrans -2- bases) epoxide) ethyoxyl) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- bridges Asia Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (300mg, 0.524mmol, 45.7% yield), it is Pale solid, LCMS (m/z):570.2[M+H]⁺。

Synthesis (4S) -7- (2- (trifluoromethyl) pyridin-4-yl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza

In room temperature by 4- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) -2- (trifluoromethyl) pyridine (9.07g, 33.2mmol) and K₃PO₄(16.27g, 77mmol) adds to the chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups of (4S) -7- Pyrido [2,3-b] [1,4] diaza(5.0g, 25.6mmol) stirring in 1,4- dioxanes (160mL) and water (40mL) In the solution mixed and the 30min that deaerates.Then in room temperature by Pd₂(dba)₃(2.340g, 2.56mmol) and X-phos (2.437g, 5.11mmol) add to reactant mixture and deaerate 5 minutes again.Then reactant mixture is stirred into 18hr at 80 DEG C.Will reaction Mixture is cooled to room temperature, and (100mL) is diluted with water, and (2X100mL) is extracted with ethyl acetate and with salt water washing (50mL).Point From organic layer, through anhydrous Na₂SO₄It is dried, filtered and concentrated (TLC eluant, eluents：100% ethyl acetate R_f0.2；UV activation). Crude compound is eluted with 80% ethyl acetate/petroleum ether, obtained through flash column chromatography (silica gel 60-120 mesh) (4S) -7- (2- (trifluoromethyl) pyridin-4-yl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine(6.5g, 20.88mmol, 82% yield), it is yellow solid, LCMS (m/z):307.0(M+H)⁺。

Synthesize (4S)-N- (4- bromopyridine -2- bases) -7- (2- (trifluoromethyl) pyridin-4-yl) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

DIPEA (7.60g, 58.8mmol) and triphosgene (5.81g, 19.59mmol) are added into (4S) -7- in 25 DEG C of priorities (2- (trifluoromethyl) pyridin-4-yl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza In the solution of (6.0g, 19.59mmol) in tetrahydrofuran (THF) (120mL), stir 1h and add 4- bromopyridine -2- amine (6.78g, 39.2mmol), then heats 18hr at 70 DEG C.Reactant mixture is cooled to 28 DEG C, then distributed in water Between (100mL) and EtOAc (100mL).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and filtrate is evaporated into (TLC Eluant, eluent：100% ethyl acetate R_f0.3；UV activation).Crude compound purifies (60-120 mesh) silica gel through column chromatography, uses 70% ethyl acetate/hexane elute), obtain (4S)-N- (4- bromopyridine -2- bases) -7- (2- (trifluoromethyl) pyridin-4-yl) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (4.5g, 8.40mmol, 42.9% yield), it is pale solid, LCMS (m/z):506.94(M+H)⁺。

Synthesize (4S)-7- (2,2- difluoros benzo [d] [1,3] Dioxol-4 -yl)-2,3,4,5- tetrahydrochysenes-1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza

By chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7-(0.6g, 10min 3.07mmol) is purged with nitrogen in the solution in the dioxane of Isosorbide-5-Nitrae-(20mL), water (4.00mL), (2,2- are then added Difluoro benzo [d] [1,3] Dioxol-4 -yl) boric acid (0.929g, 4.60mmol) and K₂CO₃(1.272g, 9.20mmol), 10min is purged with nitrogen again, then adds Pd (Ph₃P)₄(0.106g,0.092mmol).Gained reaction is mixed Compound heats 16hr at 100 DEG C.Reactant mixture is cooled to room temperature, water (50mL) is added and stirs 20min.By consolidating for acquisition Body is filtered, and is washed with ether (10mL) and pentane (5mL), is obtained (4S) -7- (2,2- difluoro benzo [d] [1,3] dioxanes Amylene -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(420mg, 1.055mmol, 34.4% yield), it is brown solid, LCMS (m/z):318.10(M+H)⁺.Synthesize the bromo- 7- of (4S) -8- chloro- 2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza

At 70 DEG C in a nitrogen atmosphere to the chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyridos [2,3-b] of (4S) -7- [1,4] diazaIn the solution of the stirring of (3g, 15.33mmol) in chloroform (20mL) add NBS (3.00g, 16.87mmol).Gained reactant mixture is stirred into 1hr at 70 DEG C.Reaction process is monitored by TLC, TLC indicates initial substance Exhaust, form nonpolar spot.Reactant mixture is cooled to RT and is diluted with water (100mL), is extracted with DCM (2x100mL). Organic layer merges and uses water (100mL), saline solution (50mL) washing, through anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain To the compound of clear yellow viscous, it is washed with pentane and fully dried, the bromo- 7- of (4S) -8- chloro- 2,3,4,5- tetra- are obtained Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(2g, 7.28mmol, 47.5% yield), it is solid for yellow Body, LCMS (m/z):276.05(M+H)⁺。

Synthesize (4S) -8- methyl -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza

By chloro- 8- methyl -2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7-(400mg, 1.908mmol), (3- (trifluoromethyl) phenyl) boric acid (290mg, 1.526mmol) and cesium carbonate (622mg, 1.908mmol) suspension in dioxane (10mL) the ＆ water of Isosorbide-5-Nitrae-(3.3mL) stirs and uses argon gas in room temperature degassing 15min, By PdCl₂(dppf)-CH₂Cl₂Adduct (1558mg, 1.908mmol) adds to reactant mixture.Then reactant mixture is existed 90 DEG C of stirring 16hr.Reactant mixture is cooled to room temperature, then filters and is washed (100ml) with EtOAc through diatomite.Draw Filtrate simultaneously concentrates and dissolved (50ml) with EtOAc.EtOAc layers are successively washed and are used in combination with water (100ml) and saline solution (100mL) Na₂SO₄Drying and dehydrating, filters and concentrates, obtain crude product.Purification of crude product, uses silica gel (100-200 mesh) and 50% EtOAc/ hexanes obtain (4S) -8- methyl -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-bridge as eluant, eluent Methylene pyridine simultaneously [2,3-b] [1,4] diaza(500mg, 1.518mmol, 80% yield) (TLC eluant, eluents：Net EtOAc: R_f-0.4.；UV activation), LCMS (m/z):320.18[M+H]⁺。

Synthesize (4S)-N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- Base) -8- methyl -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To solid (4S) -8- methyl -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diazaAdded in the solution of (500mg, 1.566mmol) in tetrahydrofuran (THF) (15mL) solid Body triphosgene (279mg, 0.939mmol), DIPEA (1.641mL, 9.39mmol) and is stirred at room temperature 30 minutes under a nitrogen. (R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- amine is then added thereto (562mg, 2.505mmol), 15h is kept 30 minutes under the conditions of seal pipe at 80 DEG C.Pass through TLC and LCMS monitoring reactions.Will Reactant mixture is diluted with water (100ml) and is extracted with ethyl acetate (2 × 100ml).The organic layer anhydrous Na of merging₂SO₄It is dry It is dry and be concentrated under reduced pressure, obtain crude product.Purified on column chromatography purify, using silica gel (100-200) and with 70%EtOAc/ oneself Alkane (gradient system) is eluted, and obtains (4S)-N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridin-4-yl) -8- methyl -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (400mg, 0.689mmol, 44.0% yield), LCMS (m/z):570.34[M+H]⁺。

Synthesize (4S)-N- (4- (2,2- dimethyl -3,6- dihydro -2H- pyrans -4- bases) pyridine -2- bases) -7- (2- methyl Pyridin-4-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

TEA (0.998mL, 7.16mmol) and triphosgene (354mg, 1.194mmol) are added in room temperature under a nitrogen (4S) -7- (tolyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(300mg, 1.194mmol) in the solution of the stirring in tetrahydrofuran (THF) (50mL).30min is stirred at room temperature in reactant mixture, Add 4- (2,2- dimethyl -3,6- dihydro -2H- pyrans -4- bases) pyridine -2- amine (731mg, 3.58mmol) and mix reaction Thing stirs 16h at 65 DEG C.Reactant mixture is cooled to room temperature, evaporating completely solvent is depressurized, then distribute in water (20mL) and Between EtOAc (50mL).Organic layer is separated, through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain crude product (4S)-N- (4- (2,2- dimethyl -3,6- dihydro -2H- pyrans -4- bases) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(220mg, 0.370mmol, 31.0% are received -5 (2H)-formamides Rate), it is brown solid, LCMS (m/z):483.26[M+H]⁺。

Synthesize chloro- 8- methyl -2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of 7-

By chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of the bromo- 7- of 8-(1g, 3.64mmol), 2,4,6- trimethyls -1,3, the boron azacyclohexane (0.457g, 3.64mmol) of 5,2,4,6- trioxa three and carbonic acid Stirred and with argon gas in room temperature in suspension of the potassium (1.510g, 10.93mmol) in 1,4- dioxanes (15mL) ＆ water (5mL) Deaerate 15min, by PdCl₂(dppf)-CH₂Cl₂Adduct (2.97g, 3.64mmol) adds to reactant mixture.Then will reaction Mixture stirs 16hr at 90 DEG C.Reactant mixture is cooled to room temperature, then filters and is washed with EtOAc through diatomite (100mL).Filtrate concentration is dissolved (100ml) with EtOAc.EtOAc layers are successively washed with water (100ml) and saline solution (50mL) Wash, use Na₂SO₄Drying and dehydrating, filters and concentrates, obtain crude product.Purified on column chromatography is purified, using neutral alumina simultaneously Eluted with 40%EtOAc/ hexanes (gradient system), obtain the chloro- 8- methyl -2,3 of desired product 7-, 4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-bridge Methylene pyridine simultaneously [2,3-b] [1,4] diaza(500mg, 2.194mmol, 60.2% yield), it is faint yellow solid, LCMS(m/z):210.11[M+H]⁺。

Synthesize (4S) -7- (3- (difluoromethyl) phenyl) -8- methyl -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza

By chloro- 8- methyl -2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7-(400mg, 1.908mmol), (3- (difluoromethyl) phenyl) boric acid (328mg, 1.908mmol) and potassium carbonate (791mg, 5.72mmol) stirred in the suspension in dioxane (15mL) the ＆ water of Isosorbide-5-Nitrae-(4mL) and with argon gas in room temperature degassing 15min, will PdCl₂(dppf)-CH₂Cl₂Adduct (1558mg, 1.908mmol) adds to reactant mixture.Then by reactant mixture 90 DEG C stirring 16hr.Reactant mixture is cooled to room temperature, then filters and is washed (100ml) with EtOAc through diatomite.By filtrate Concentration is dissolved (50ml) with EtOAc.EtOAc layers are successively washed with water (50ml) and saline solution (50mL) and use Na₂SO₄Dry Dehydration, filters and concentrates, obtain crude product.Purified on column chromatography is purified, and using silica gel (100-200) and uses 50%EtOAc/ Hexane (gradient system) is eluted, and obtains desired product (4S) -7- (3- (difluoromethyl) phenyl) -8- methyl -2,3,4,5- tetra- Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(300mg, 0.969mmol, 50.8% yield), it is light Yellow solid LCMS (m/z):302.11[M+H]⁺。

Synthesis (4S) -7- (3- (difluoromethyl) phenyl)-N- (2- (((R) -2,2- dimethyl -1,3- dioxolane - 4- yls) methoxyl group) pyridin-4-yl) -8- methyl -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

To solid (4S) -7- (3- (difluoromethyl) phenyl) -8- methyl -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diazaAdded in the solution of (250mg, 0.830mmol) in tetrahydrofuran (THF) (15mL) solid Body triphosgene (148mg, 0.498mmol), DIPEA (0.869mL, 4.98mmol) and is stirred at room temperature 30 minutes under a nitrogen. (R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- amine is then added thereto (298mg, 1.327mmol), 15h is kept 30 minutes under the conditions of seal pipe at 75 DEG C.Pass through TLC and LCMS monitoring reactions.Will Reactant mixture is diluted with water (10ml) and is extracted with ethyl acetate (2 × 50ml).The organic layer anhydrous Na of merging₂SO₄Dry And be concentrated under reduced pressure, obtain crude product, LCMS (m/z):551.91[M+H]⁺。

Synthesize (4S)-N- (4- bromopyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to solid (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- bridges under nitrogen gas stirring Methylene pyridine simultaneously [2,3-b] [1,4] diazaThe solution of (2.5g, 9.91mmol) in tetrahydrofuran (THF) (40mL) Middle addition Solid triphosgene (1.764g, 5.94mmol), is stirred at room temperature 30 minutes under a nitrogen.DIPEA is added thereto (8.65mL, 49.5mmol) and 4- bromopyridine -2- amine (2.57g, 14.86mmol).Reactant mixture is stirred into 16hr at 65 DEG C. Monitored and reacted by TLC.It is added to organic in ethyl acetate and is washed with water 50mL and saturated brine 100mL, through Na₂SO₄It is dry It is dry and be evaporated in vacuo, crude product is obtained, it is brown solid, crude product is added in neutral alumina and eluted with EtOAc, collected Fraction obtains compound, is washed with ether and pentane, then filters and is washed with ether, obtains pure compound (4S)-N- (4- Bromopyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (1.72g, 2.97mmol, 30.0% yield), LCMS (m/z):453.20[M+H]⁺。

Synthesize (4S)-N- (5- bromopyridine -3- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diazaTriphosgene is added in the solution of the stirring of (8g, 31.7mmol) in tetrahydrofuran (THF) (130mL) (5.65g, 19.02mmol), DIPEA (27.7mL, 159mmol).It is stirred at room temperature under a nitrogen 30 minutes.Then add thereto Enter 5- bromopyridine -3- amine (8.23g, 47.6mmol), 16h is kept at 75 DEG C under the conditions of seal pipe.Monitored by TLC and LCMS Reaction.By reactant mixture distribution between ethyl acetate (200mL) and water (100mL).Organic solution water and salt water washing, Through Na₂SO₄It is dried, filtered and concentrated, obtains crude compound.Crude product is added in neutral alumina column, then with 50% EtOAc/ petroleum ethers are eluted.Collect fraction and concentrate, obtain compound and washed with pentane, obtain pure compound (4S)-N- (5- bromopyridine -3- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine- 5 (2H)-formamides (6.2g, 12.72mmol, 40.1% yield), it is white solid, LCMS (m/z):451.19 [M+H]⁺。

Synthesize (4S)-N- (the bromo- 1- methyl -2- oxos -1,2- dihydropyridines -3- bases of 5-) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In a nitrogen atmosphere to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diazaAdd in the solution of the stirring of (1.0g, 3.96mmol) in tetrahydrofuran (THF) (30mL) Enter TEA (2.76mL, 19.82mmol) and triphosgene (1.176g, 3.96mmol).Gained reactant mixture is stirred at room temperature 1hr.Bromo- 1- picolines -2 (1H) -one (0.805g, 3.96mmol) of 3- amino -5- are added into reactant mixture, then 70 DEG C stirring 16hr.Reaction process is monitored by TLC.(50mL) is diluted with water in reactant mixture, and (3X50mL) is extracted with EtOAc, Organic layer merges and through anhydrous Na₂SO₄Dry, filter and be concentrated in vacuo, obtain crude compound.Thick material is purified through column chromatography (100-200 mesh silica gel, eluted with 2%MeOH/DCM), obtains (4S)-N- (bromo- 1- methyl -2- oxos -1,2- dihydro pyrroles of 5- Pyridine -3- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (800mg, 1.338mmol, 33.8% yield), are brown solid compound, LCMS (m/z):481.22[M+ H]⁺。

Synthesize (4S) -7- (1- ethyl -1H- pyrazoles -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza

In room temperature to the chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyridos [2,3-b] of (4S) -7- under nitrogen gas stirring [1,4] diaza(3g, 15.33mmol), 1- ethyls -4- (4,4,5,5- tetramethyls -1,3,2- dioxaborolans -2- Base) phosphoric acid is added in the solution of -1H- pyrazoles (4.09g, 18.40mmol) in 1,4- dioxanes (40mL), water (10.00mL) Tripotassium (9.76g, 46.0mmol).Reactive material is deaerated 15min with nitrogen.PdCl is added thereto in room temperature₂(dppf)- CH₂Cl₂Adduct (1.252g, 1.533mmol) and the 15min that deaerates again.Reactant mixture is stirred into 16hr at 100 DEG C.Pass through TLC monitors reaction process.TLC indicates that initiation material exhausts, and forms new spot.Reactive material is cooled down to room temperature, acetic acid second is used Ester (100mL) dilutes, and is filtered through diatomite, collects filtrate, and washed with water (100mL).Organic layer is through Na₂SO₄Dry, mistake Filter and concentrate, obtain crude compound.Crude product adds to silica gel (100-200) post and uses DCM/MeOH, and 5%MeOH/DCM is washed It is de-.Enriched product fraction, obtains (4S) -7- (1- ethyl -1H- pyrazoles -4- bases) -2,3,4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-endo-methylene group pyrrole Pyridine simultaneously [2,3-b] [1,4] diaza(2g, 7.83mmol, 51.1% yield), it is green solid, LCMS (m/z): 256.22(M+H)⁺。

Synthesize (4S) -8- chloro- 7- (1- ethyl -1H- pyrazoles -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza

In room temperature to (4S) -7- (1- ethyl -1H- pyrazoles -4- bases) -2,3,4,5- tetrahydrochysene -1,4- bridges under nitrogen gas stirring Methylene pyridine simultaneously [2,3-b] [1,4] diazaNCS is added in the solution of (2g, 7.83mmol) in chloroform (20mL) (1.255g,9.40mmol).1hr is stirred at room temperature in reactant mixture.Reaction process is monitored by TLC.TLC indicates that starting is former Material exhausts, and forms new spot, R_fFor 0.3.Water (50mL) is added into reactant mixture and extracted (50mL) with DCM.Organic layer Through Na₂SO₄It is dried, filtered and concentrated, obtains thick material, it is the compound of viscous brown.Crude product adds to silica gel (100- 200) post and CH is used₂Cl₂/ MeOH, 3% methanol/DCM elutions, eluted product.Enriched product fraction, obtains the chloro- 7- of (4S) -8- (1- ethyl -1H- pyrazoles -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza (1.2g, 3.15mmol, 40.2% yield), it is the compound of light brown viscous, LCMS (m/z):290.04(M+H)⁺。

Synthesize (4S) -8- chloro- N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -4- bases) -7- (1- ethyl -1H- pyrazoles -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides

In room temperature to the chloro- 7- of (4S) -8- (1- ethyl -1H- pyrazoles -4- bases) -2,3,4,5- tetrahydrochysene -1 under nitrogen gas stirring, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(500mg, 1.726mmol) is in tetrahydrofuran (THF) (20mL) Solution in add TEA (1.203mL, 8.63mmol), triphosgene (512mg, 1.726mmol).By reactant mixture in room temperature Stir 30min.(R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- amine is added thereto (387mg,1.726mmol).In room temperature, reactant mixture is stirred into 6hr at 80 DEG C.Reaction process is monitored by TLC.TLC refers to Show that initiation material exhausts, form new spot, R_fFor 0.4R_f.Reactive material is cooled down to room temperature, (100mL) is diluted with water and second is used Acetoacetic ester extracts (100mL).Organic layer is through anhydrous Na₂SO₄It is dried, filtered and concentrated, obtains thick material, it is viscous brown Compound.Crude product is added into combiflash silica gel (40g) post and eluted with 3%DCM/MeOH.The pure product fraction of concentration, Obtain the chloro- N- of (4S) -8- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridin-4-yl) - 7- (1- ethyl -1H- pyrazoles -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (500mg, 0.881mmol, 51.1% yield) (N37522-56-A2), it is pale solid, LCMS (m/ z):539.95(M+H)⁺。

Synthesize chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7,8- two

At 0 DEG C to chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7- NCS (5.12g, 38.3mmol) is added portionwise in the solution of the stirring of (5g, 25.6mmol) in chloroform (50mL), and at 26 DEG C Stir 6hr.Reactant mixture is concentrated in vacuo and by residue distribution between water (50mL) and DCM (2X100mL).Separation has Machine layer and through anhydrous Na₂SO₄Dry, filter and simultaneously evaporate filtrate, obtain pure (4S) -7,8- bis- chloro- 2,3,4,5- tetrahydrochysene -1, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(5g, 18.83mmol, 73.7% yield), it is faint yellow solid, LCMS(m/z):230.02[M+H]

Synthesize (4S) -8- chloro- N- (2- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -4- bases) -7- (1- ethyl -1H- pyrazoles -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides

In room temperature to the chloro- 7- of (4S) -8- (1- ethyl -1H- pyrazoles -4- bases) -2,3,4,5- tetrahydrochysene -1 under nitrogen gas stirring, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(500mg, 1.726mmol) is in tetrahydrofuran (THF) (30mL) Solution in add TEA (1.203mL, 8.63mmol), triphosgene (512mg, 1.726mmol).By reactant mixture in room temperature Stir 30min.(S) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrrole is added thereto in room temperature Pyridine -4- amine (387mg, 1.726mmol).Reactant mixture is stirred into 6hr at 80 DEG C.Reaction process is monitored by TLC.TLC refers to Show that initiation material is depleted.Reactant mixture is cooled down to room temperature, (50mL) is diluted with water, is extracted with ethyl acetate (100mL).Have Machine layer is through anhydrous Na₂SO₄It is dried, filtered and concentrated, obtains thick material, it is the compound of viscous brown.Crude product is added to Combiflash silica gel (40g) post is simultaneously eluted with DCM/MeOH, 2% methanol/DCM.Eluted product.Enriched product fraction, is obtained The chloro- N- of (4S) -8- (2- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridin-4-yl) -7- (1- Ethyl -1H- pyrazoles -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine (450mg, 0.802mmol, 46.5% yield), LCMS (m/z):540.30(M+H)⁺。

Synthesize iodo- 3,4- dihydros -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of the chloro- 8- of (4S) -7--5 (2H)-t-butyl formate

To iodo- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of the chloro- 8- of (4S) -7- Boc is added in the suspension of (31g, 96mmol) in tetrahydrofuran (THF) (300mL)₂O (33.6mL, 145mmol) and DMAP (14.13g,116mmol).Then reactant mixture is stirred into 20hr at 65 DEG C.Monitored and reacted by TLC.Reactive material water Dilution (100mL) is simultaneously extracted twice (2 × 250mL) with EtOAc.The organic layer aqueous salt solu-tion of merging simultaneously uses Na₂SO₄It is dry Dry dehydration, filters and evaporates and obtain crude product.Crude product carries out chromatography on neutral alumina, is washed with 3% ethyl acetate/petroleum ether It is de-, obtain iodo- 3,4- dihydros-Isosorbide-5-Nitrae-endo-methylene group pyrido [2,3-b] [Isosorbide-5-Nitrae] diazas of the pure chloro- 8- of compound (4S) -7-- 5 (2H)-t-butyl formates (26.8g, 51.3mmol, 53.2% yield), LCMS (m/z):422.00(M+H)⁺。

Synthesize iodo- 8- methyl -3,4- dihydros -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazines of the chloro- 9- of (4S) -7- It is miscellaneous- 5 (2H)-t-butyl formates

LDA (23.72mL, 47.4mmol) is added into the iodo- 3,4- bis- of the chloro- 8- of (4S) -7- at -78 DEG C under nitrogen gas stirring Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-t-butyl formates (10.0g, 23.72mmol) exist In the solution of stirring in tetrahydrofuran (THF) (100mL).Then reactant mixture is stirred 30 minutes at -78 DEG C.Then Iodomethane (2.97mL, 47.4mmol) is added into reactant mixture at -78 DEG C.Then reactant mixture is warmed to room temperature.So 16hr is stirred at room temperature in reactant mixture afterwards.Monitored and reacted by TLC.By reactant mixture saturation NH₄Cl solution is quenched And extracted with EtOAc.EtOAc layers of priority water and aqueous salt solu-tion simultaneously use Na₂SO₄Drying and dehydrating, filters and concentrates, obtain Crude product.Crude product adds in neutral alumina and uses 1%EtOAc/ Hex.The fraction of collection is evaporated, obtain (4S)- Iodo- 8- methyl -3,4- dihydros -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of the chloro- 9- of 7-- 5 (2H)-formic acid uncles Butyl ester (4.0g, 8.07mmol, 34.0% yield), it is pale solid, LCMS (m/z):436.04(M+H)⁺。

Synthesize the iodo- 8- methyl -2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyridos [2,3-b] [1,4] of the chloro- 9- of (4S) -7- Diaza

Ether/HCl is added into the iodo- 8- methyl -3,4- dihydros -1,4- bridge methylenes of the chloro- 9- of (4S) -7- at 0 DEG C under a nitrogen Yl pyridines simultaneously [2,3-b] [1,4] diazaIn the solution of stirring in -5 (2H)-t-butyl formates (1.5g, 3.44mmol). Reactant mixture is stirred into 6hr at 26 DEG C.Monitored and reacted by TLC.Reactive material is filtered, solid is taken out, uses saturation NaHCO₃ Solution (50mL) is quenched and extracted (100ml) with DCM.DCM layers of priority water and aqueous salt solu-tion simultaneously use Na₂SO₄Dry de- Water, filters and concentrates, and obtains the iodo- 8- methyl -2,3 of the chloro- 9- of thick material (4S) -7-, 4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-endo-methylene group pyrido [2,3-b] [1,4] diaza(1.0g, 2.55mmol, 74.2% yield), it is white solid, LCMS (m/z):336.8(M +H)⁺。

Synthesize the chloro- 8,9- dimethyl -2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyridos [2,3-b] [1,4] of (4S) -7- Diaza

By the iodo- 8- methyl -2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two of the chloro- 9- of (4S) -7- AzepineThe boron azacyclohexane (Trimethylboroxine) of (2.5g, 7.45mmol), trimethyl trioxa three (2.483mL, 7.45mmol) stirred simultaneously with suspension of the potassium carbonate (3.09g, 22.35mmol) in 1,4- dioxanes (50mL) ＆ water (5mL) Deaerated 15 minutes in room temperature with argon gas, tetrakis triphenylphosphine palladium (0) (0.861g, 0.745mmol) is added into reactant mixture.So Reactant mixture is stirred into 16hr at 90 DEG C afterwards.Monitored and reacted by TLC.Reactant mixture is cooled to room temperature, then through silicon Diatomaceous earth is filtered and washed with EtOAc.Concentration filtrate is simultaneously dissolved with EtOAc.EtOAc layers are successively used in combination with water and aqueous salt solu-tion Na₂SO₄Drying and dehydrating, filters and concentrates, obtain crude product.Crude product adds in neutral alumina and uses 10%EtOAc/ hexanes Elution.The fraction of collection is evaporated, chloro- 8, the 9- dimethyl -2,3 of (4S) -7-, 4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-endo-methylene group pyridine is obtained And [2,3-b] [1,4] diaza(1.2g, 5.08mmol, 68.2% yield), it is faint yellow solid, LCMS (m/z): 224.08(M+H)⁺。

Synthesize (4S) -8,9- dimethyl -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza

By the chloro- 8,9- dimethyl -2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two of (4S) -7- Azepine(1.200g, 5.36mmol), (2- picoline -4- bases) boric acid (0.918g, 6.71mmol) and tripotassium phosphate The suspension of (3.42g, 16.09mmol) in 1,4- dioxanes (20mL) ＆ water (3.0mL) stirs and uses argon gas to be deaerated in room temperature 15 minutes.Then by Pd₂(dba)₃(0.246g, 0.268mmol) and X-Phos (0.256g, 0.536mmol) add to reaction mixing In thing.Then reactant mixture is stirred into 16hr at 90 DEG C.Monitored and reacted by TLC.Reactant mixture is cooled to room temperature, so Filter and washed with EtOAc by diatomite.Draw filtrate and concentrate and dissolved with EtOAc.EtOAc layers of priority water and salt solution Solution washs and uses Na₂SO₄Drying and dehydrating, filters and concentrates, obtain crude product.Crude product adds in neutral alumina and uses DCM Elution.The fraction of collection is evaporated, (4S) -8,9- dimethyl -7- (2- picoline -4- bases) -2 is obtained, 3,4,5- tetrahydrochysene -1, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(0.700g, 2.495mmol, 46.5% yield), it is faint yellow Solid.LCMS(m/z):281.30(M+H)⁺。

Synthesis (4S) -7- (1- cyclopropyl -1H- pyrazoles -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza

In room temperature by tripotassium phosphate (6.51g, 30.7mmol) and 1- cyclopropyl -4- (4,4,5,5- tetramethyls -1,3,2- two The amyl- 2- yls of oxa- boron heterocycle) -1H- pyrazoles (2.87g, 12.27mmol) adds to the chloro- 2,3,4,5- tetrahydrochysenes -1,4- bridges of (4S) -7- Methylene pyridine simultaneously [2,3-b] [1,4] diaza(2g, 10.22mmol) is in 1,4- dioxanes (40mL), water (10.00mL) Mixture in stirring solution in.Purification for argon 5min is used, PdCl is then added₂(dppf)-CH₂Cl₂Adduct (0.835g, 1.022mmol), in 110 DEG C of stirring reaction mixtures, keeps 16h.Reactant mixture is reached RT, be diluted with water (150mL) and it is extracted with ethyl acetate (2x300mL), with salt water washing (200mL).The organic layer of merging is through Na₂SO₄Dry simultaneously It is concentrated under reduced pressure, obtains crude compound.Crude product is through flash column chromatography (silica gel：100-200 mesh) and use 10%MeOH- DCM is eluted, and obtains (4S) -7- (1- cyclopropyl -1H- pyrazoles -4- bases) -2,3,4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-endo-methylene group pyrido [2, 3-b] [1,4] diaza(1g, 3.52mmol, 34.4% yield), LCMS (m/z):268.13[M+H]⁺。

Synthesize (4S) -8- chloro- 7- (1- cyclopropyl -1H- pyrazoles -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza

By (4S) -7- (1- cyclopropyl -1H- pyrazoles -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza(1g, 3.74mmol) is dissolved in chloroform (100mL), and NCS is added at 0 DEG C under nitrogen gas stirring (0.500g,3.74mmol).20min is stirred at room temperature in reactant mixture.So that reactant mixture reaches room temperature and uses 90ml Water quenching is gone out and uses 2x150mL DCM to extract.The organic layer of merging is through Na₂SO₄Dry and be concentrated under reduced pressure, obtain thick material.Crude product Through flash column chromatography (silica gel：100-200 mesh) and eluted with 10%MeOH-DCM, obtain (4S) -8- chloro- 7- (1- rings third Base -1H- pyrazoles -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(850mg, 2.54mmol, 67.8% yield), LCMS (m/z):302.42[M+H]⁺。

Synthesize (4S) -8- chloro- 7- (1- cyclopropyl -1H- pyrazoles -4- bases)-N- (2- (((S) -2,2- dimethyl -1,3- two Tetrahydrofuran -4- bases) methoxyl group) pyridin-4-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides

By the chloro- 7- of (4S) -8- (1- cyclopropyl -1H- pyrazoles -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza(400mg, 1.325mmol) is dissolved in tetrahydrofuran (THF) (20mL), under nitrogen gas stirring Triphosgene (393mg, 1.325mmol), TEA (0.924mL, 6.63mmol) are added at 0 DEG C.Reactant mixture is stirred at room temperature 30min.(S) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- amine is added thereto (446mg, 1.988mmol) and stir 16h at 80 DEG C in seal pipe.So that reactant mixture reaches room temperature and uses 50ml water quenchings Go out and use 2x150ml ethyl acetate to extract.The organic layer of merging is through Na₂SO₄Dry and be concentrated under reduced pressure, obtain crude product.Crude product Through flash column chromatography (silica gel：100-200 mesh) and eluted with 1%MeOH-DCM, obtain (4S) -8- chloro- 7- (1- rings third Base -1H- pyrazoles -4- bases)-N- (2- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (285mg, 0.510mmol, 38.4% yield), it is pale solid, LCMS (m/z):552.50[M+H]⁺。

Synthesize (4S)-N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- Base) -9- methyl -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -9- methyl -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diazaIn the solution of (450mg, 1.409mmol) in THF (30mL) add triphosgene (251mg, 0.846mmol).Then add TEA (0.196mL, 1.409mmol), then stir to room temperature, continue 1h, be subsequently added (R)- 2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- amine (474mg, 2.114mmol), at 75 DEG C Keep 16h.Pass through TLC and LCMS monitoring reactions.Reactant mixture is poured into saturation NaHCO₃Solution (30mL) simultaneously uses acetic acid second Ester extracts (2x100mL).Organic layer is through anhydrous Na₂SO₄Dry and be concentrated under reduced pressure, obtain crude product.Crude product adds to neutral alumina Aluminium post is simultaneously eluted with 20% ethyl acetate/petroleum ether.The fraction of collection is evaporated, (4S)-N- (2- (((R) -2,2- diformazans are obtained Base -1,3- dioxolane -4- bases) methoxyl group) pyridin-4-yl) -9- methyl -7- (3- (trifluoromethyl) phenyl) -3,4- two Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamide (400mg, 0.690mmol, 48.9% Yield), it is white solid, LCMS (m/z):569.93[M+H]⁺。

Synthesize (4S) -9- methyl -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza

To chloro- 9- methyl -2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7-(600mg, 2.86mmol), (3- (trifluoromethyl) phenyl) boric acid (815mg, 4.29mmol) and K₃PO₄(1822mg, 8.58mmol) in the solution of the degassing in 1,4- dioxanes (40mL), water (10mL) add x-phos (273mg, 0.572mmol)、Pd₂(dba)₃(262mg,0.286mmol).Reactant mixture is stirred into 3hr at 110 DEG C.It is anti-by TLC monitorings Should.Reactant mixture is poured into ice (50mL) and is extracted with ethyl acetate (3x100mL).Organic layer is through anhydrous Na₂SO₄Dry, Filter and be concentrated under reduced pressure, obtain crude product, neutral alumina column will be added to and eluted with 40% ethyl acetate/petroleum ether.It will receive The fraction evaporation of collection, obtains (4S) -9- methyl -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-endo-methylene group pyrrole Pyridine simultaneously [2,3-b] [1,4] diaza(600mg, 1.691mmol, 59.1% yield), it is faint yellow solid, LCMS (m/ z):320.13[M+H]⁺。

Synthesize (4S) -9- methyl -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza

To chloro- 9- methyl -2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7-(500mg, 2.385mmol), (2- picoline -4- bases) boric acid (490mg, 3.58mmol) and K₃PO₄(1519mg, 7.15mmol) in the solution of the degassing in the dioxane of Isosorbide-5-Nitrae-(40mL), water (10mL) add x-phos (227mg, 0.477mmol)、Pd₂(dba)₃(218mg,0.238mmol).Reactant mixture is stirred into 3hr at 110 DEG C.It is anti-by TLC monitorings Should.Reactant mixture is poured into ice (50mL) and is extracted with ethyl acetate (3x100mL).Organic layer is through anhydrous Na₂SO₄Dry, Filter and be concentrated under reduced pressure, obtain crude product, added to neutral alumina column and eluted with 40% ethyl acetate/petroleum ether.Will The fraction evaporation of collection, obtains (4S) -9- methyl -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-endo-methylene group Pyrido [2,3-b] [1,4] diaza(450mg, 1.653mmol, 69.3% yield), it is faint yellow solid LCMS (m/ z)267.21[M+H]⁺。

Synthesize trifluoromethanesulfonic acid 2,2- dimethyl -3,6- dihydro -2H- pyrans -4- base esters

LDA (2.75mL, 20.29mmol) is added into (3H) -one of 2,2- dimethyl dihydro -2H- pyrans -4 at -78 DEG C In the solution of the stirring of (2.0g, 15.60mmol) in tetrahydrofuran (THF) (40mL) and stirring 20 minutes.In -78 DEG C of additions 1,1,1- tri- fluoro- N- phenyl-N- ((trifluoromethyl) sulfonyl) Methanesulfomide (3.98mL, 18.73mmol), is then stirred at 28 DEG C Mix 19hr.Reactant mixture is quenched with saturated sodium bicarbonate solution, and (10mL) is diluted with water, and is extracted with ether (2X30mL), is used Aqueous salt solu-tion (20mL).Organic layer is separated, through Na₂SO₄It is dried, filtered and concentrated, obtains crude product.Crude product is through post color Spectrum purifying, uses (100-200) silica gel chromatography, and is eluted with 15%EtOAc/ hexanes (gradient system), is expected Product trifluoromethanesulfonic acid 2,2- dimethyl -3,6- dihydro -2H- pyrans -4- base esters (2.4g, 5.98mmol, 38.3% yield), It is weak yellow liquid.GCMS(m/z):260[M+H]⁺。

Synthesize 4- (2,2- dimethyl -3,6- dihydro -2H- pyrans -4- bases) pyridine -2- amine

By 4- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) pyridine -2- amine (1.0g, 4.54mmol), trifluoromethanesulfonic acid 2,2- dimethyl -3,6- dihydros -2H- pyrans -4- base esters (2.365g, 9.09mmol) and phosphoric acid Suspension of the tripotassium (2.89g, 13.63mmol) in 1,4- dioxanes (30mL) stirs and uses argon gas to be deaerated 15 points in room temperature Clock.By PdCl₂(dppf)-CH₂Cl₂Adduct (0.186g, 0.227mmol) adds to reactant mixture.Then by reactant mixture 4hr is stirred at 90 DEG C.Monitored and reacted by TLC.Reactant mixture is cooled to room temperature, then filters and is used in combination through diatomite EtOAc washs (20ml).Draw filtrate and concentrate and dissolved (20ml) with EtOAc.EtOAc layers successively with water (10ml) and salt solution Solution (10mL) is washed, and uses Na₂SO₄Drying and dehydrating, filters and concentrates, obtain crude product.Purified on column chromatography is purified, and is used Neutral alumina is simultaneously eluted with 100%DCM (gradient system), obtains desired product 4- (2,2- dimethyl -3,6- dihydro -2H- Pyrans -4- bases) pyridine -2- amine (0.8g, 3.79mmol, 83% yield), it is brown solid, LCMS (m/z):205.2[M+H ]⁺。

Synthesize 4- (1H-1,2,3- triazole -5- bases) pyridine -2- amine

In microwave 150 DEG C to 4- ethynyl pyridine -2- amine (350mg, 2.96mmol) TMSN3 (0.5mL, In the suspension of stirring in 3.77mmol).Gained reactant mixture is stirred into 30min in identical temperature.Monitored by TLC Reaction process.Reactant mixture filter, solid chemical compound wash with DCM (2mL) and drying, obtain 4- (1H-1,2,3- triazoles- 5- yls) pyridine -2- amine (360mg, 1.832mmol, 61.8% yield) light yellow solid, LCMS (m/z):162.1[M+H]⁺。

Synthesize 3- (PA -4- base) oxazolidine -2- ketone

In the solution of stirring of the room temperature to 4- bromopyridine -2- amine (1g, 5.78mmol) in 1,4- dioxanes (10ml) Jia Ru oxazolidine -2- ketone (0.755g, 8.67mmol), potassium carbonate (1.598g, 11.56mmol), cuprous iodide (I) (0.110g, 0.578mmol), N, N'- dimethyl-ethylenediamine (0.102g, 1.156mmol).By gained reactant mixture in microwave 110 DEG C stirring 1hr.Reaction process is monitored by TLC.The solvent of reactant mixture is evaporated, (10mL) and 20 points of stirring is diluted with water Clock, forms solid, filters and dries, acquisition 3- (PA -4- base) oxazolidine -2- ketone (580mg, 3.11mmol, 53.8% yield), it is faint yellow solid, LCMS (m/z):180.0[M+H]⁺。

Synthesize 4- (1H-1,2,3- triazol-1-yls) pyridine -2- amine and 4- (2H-1,2,3- triazole -2- bases) pyridine -2- amine

In room temperature to 1H-1,2,3- triazoles (0.599g, 8.67mmol), 4- bromopyridine -2- amine (1g, 5.78mmol) 1, K is added in solution in 4- dioxanes (10mL)₂CO₃(1.598g, 11.56mmol), cuprous iodide (I) (0.110g, 0.578mmol) and N, N- dimethyl-ethylenediamine (0.126mL, 1.156mmol).Reactant mixture is stirred in microwave at 110 DEG C Mix 1hr.Reactant mixture is quenched with water (30mL), and successively extracted with ethyl acetate (2 × 50mL) and saline solution (50mL) Take, separate each layer, use anhydrous Na₂SO₄It is dried, filtered and concentrated, obtains crude product.Purification of crude product, obtains 4- (1H-1,2,3- Triazol-1-yl) pyridine -2- amine (0.1g, 0.562mmol, 9.72% yield), it is pale solid, and 4- (2H-1,2,3- Triazole -2- bases) pyridine -2- amine (0.3g, 1.787mmol, 30.9% yield), it is pale solid, LCMS (m/z):162.0 [M+H]⁺。

Synthesize (E) -2- amino-N- ((dimethylamino) methine) Pyrazinamide

In room temperature to PA -4- formamides (5g, 36.5mmol) at N,N-dimethylformamide (DMF) (50mL) In stirring solution in add 1,1- dimethoxys-N, N- dimethyl methylamine (9.76mL, 72.9mmol).Gained reaction is mixed Compound stirs 4hr at 27 DEG C.Reaction process is monitored by TLC.The solvent of reactant mixture is evaporated, with EtOAc (5X20mL) With icy water dilution, the organic layer and vacuum concentration of separation obtain (E) -2- amino-N- ((dimethylamino) methine) pyrrole Pyridine -4- formamides (4.5g, 23.41mmol, 64.2% yield), it is pale solid, LCMS (m/z):193.01[M+H]⁺。

Synthesize 4- (1H-1,2,4- triazole -5- bases) pyridine -2- amine

In room temperature to (E) -2- amino-N- ((dimethylamino) methine) Pyrazinamide (4g, 20.81mmol) in ethanol In stirring in (40mL) suspension in add hydrazine hydrate (1.042g, 20.81mmol).By gained reactant mixture identical Temperature stirring 16hr.Reaction process is monitored by TLC.Reactant mixture is filtered, and solid chemical compound is washed (20mL) with EtOAc And dry filtrate, obtain 2g (LCMS shows that 40% is desired product) solid chemical compound.By compound through preparation HPLC Purifying, obtains 4- (1H-1,2,4- triazole -5- bases) pyridine -2- amine (750mg, 4.40mmol, 21.13% yield), it is brown Solid, LCMS (m/z):162.0[M+H]⁺。

Synthesize 6- (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethyoxyl) pyridine -2- amine

0 DEG C by 2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethanol (1.5g, 10.26mmol) in N- methyl -2- pyrroles Solution in alkanone (NMP) (4mL) is added in NaH (0.616g, 25.7mmol) suspension, and by reactant mixture at 28 DEG C Stir 30min.6- fluorine pyridine -2- amine (1.150g, 10.26mmol) is added into reactant mixture at 0 DEG C, and by reactant mixture 16hr is stirred at 120 DEG C.Reactant mixture is quenched with cold water and uses dichloromethane (2 × 60mL) to extract.Organic layer water (30mL) and saturated brine solution (20mL) are washed, through anhydrous Na₂SO₄It is dried, filtered and concentrated.By thick material through neutral alumina Aluminium purification by flash chromatography.Thick material is diluted with DCM, is then absorbed with neutral alumina and uses 20-25-%EtOAc/ petroleum ethers Elution, collect fraction simultaneously concentrate, obtain 6- (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethyoxyl) pyridine -2- amine (700mg, 2.66mmol, 25.9% yield), LCMS (m/z):239.0[M+H]⁺。

Synthesize 3- ((6- aminopyridine -2- bases) epoxide) propyl- 1- alcohol

0 DEG C by propyl- 1,3- glycol (1.358g, 17.84mmol) add to NaH (1.070g, 44.6mmol) N- methyl- In the solution of stirring in 2-Pyrrolidone (NMP) (5mL) and stirring 1h, be subsequently added into 6- fluorine pyridine -2- amine (1.0g, 8.92mmol) and at 80 DEG C stir 2h.Reactive material is cooled to room temperature, is added slowly in icy water and dilute with ethyl acetate Release.The organic layer water and salt water washing of separation.Organic layer is through Na₂SO₄It is dried, filtered and concentrated, obtains crude compound.Slightly Produced compounds are eluted using 100-200 silica gel purifications and in 100% ethyl acetate, obtain 3- ((6- aminopyridine -2- bases) oxygen Base) propyl- 1- alcohol (0.4g, 1.760mmol, 19.73% yield), it is viscous brown thing, LCMS (m/z):169.22[M+H]⁺。

Synthesize 6- (3- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) propoxyl group) pyridine -2- amine

At 0 DEG C by 3- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) propyl- 1- alcohol (4.2g, 26.2mmol)/1,4- dioxanes (20mL) is added in solution of the NaH (1.307g, 32.7mmol) in the dioxane of Isosorbide-5-Nitrae-(20mL), and by reactant mixture 28 DEG C stirring 30min.6- chloropyridine -2- amine (2.8g, 21.78mmol)/1,4- dioxanes (20mL) are added into reaction mixing at 0 DEG C Thing, and reactant mixture is stirred into 10hr at 100 DEG C.By reactant mixture distribution water (20mL) and DCM (2 × 25mL) it Between.DCM layers are used saturation NaHCO₃Solution is washed, and is separated and through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain rough 6- (3- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) propoxyl group) pyridine -2- amine (5.5g, 18.12mmol, 83% yield), its For brown oil, LCMS (m/z):253.2[M+H]⁺。

Synthesize 3- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) propyl- 1- alcohol

0 DEG C by p-methyl benzenesulfonic acid monohydrate (0.678g, 3.57mmol) add to propyl- 1,3- glycol (5.43g, 71.3mmol) with the solution of stirring of the 3,4- dihydro -2H- pyrans (3g, 35.7mmol) in dichloromethane (DCM) (50mL) In.Reactant mixture is stirred into 2h at 28 DEG C.By reactant mixture distribution between water (20mL) and DCM (2 × 25mL).DCM Layer uses saturation NaHCO₃Solution is washed, and is separated and through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain rough 3- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) propyl- 1- alcohol (4.2g, 26.2mmol, 73.5% yield), it is colorless oil.

Synthesize 3- ((6- aminopyridine -2- bases) epoxide) -2,2- dimethyl propylene -1- alcohol

2,2- dimethyl propylene -1,3- glycol (3.0g, 28.8mmol) is added into NaH (2.304g, 57.6mmol) at 0 DEG C to exist In the solution of stirring in METHYLPYRROLIDONE (NMP) (25mL).30min is stirred at room temperature in reactant mixture.0 DEG C 6- chloropyridine -2- amine (4.44g, 34.6mmol) is added into reactant mixture.Reactant mixture is stirred into 16h at 100 DEG C, and Reaction process is monitored by TLC.Reactant mixture is cooled to room temperature, is quenched and is extracted with ethyl acetate with icy water (3X30mL).Organic layer is washed with water (30mL) and saturated brine solution (30mL), through anhydrous Na₂SO₄It is dried, filtered and concentrated, Obtain crude compound, TLC eluant, eluents：50%EtOAc/ hexanes, R_f:0.3, UV activation.Crude compound is pure through column chromatography Change, eluted using neutral alumina and with 5%EtOAc/ petroleum ethers, obtain pure 3- ((6- aminopyridine -2- bases) epoxide) -2, 2- dimethyl propylene -1- alcohol (1.5g, 6.79mmol, 23.56% yield), it is pale solid.LCMS:(m/z):197.16[M +H]⁺。

Synthesize 4- ((6- aminopyridine -2- bases) epoxide) -2- methyl butyl- 2- alcohol

At 0 DEG C.To under a nitrogen in 0 DEG C of NaH (1.167g, 29.2mmol) stirred in METHYLPYRROLIDONE (NMP) 3- methyl butyl- 1,3- glycol (3.04g, 29.2mmol) is added dropwise in the suspension in (2mL) in N- methyl -2- pyrrolidines In solution in ketone (NMP) (2mL), continue 10min.After 10min, be added dropwise at 0 DEG C 6- chloropyridine -2- amine (2.5g, 19.45mmol) the solution in METHYLPYRROLIDONE (NMP) (2mL), continues 10min.By reactant mixture at 120 DEG C Heat 16hr.Reaction process is monitored through TLC.Reactant mixture is poured into frozen water and (3X30ml), organic solvent are extracted with EtOAc Through Na₂SO₄Dry and be concentrated in vacuo, obtain thick material.Thick material is purified through column chromatography, using silica gel (100-200 mesh), uses 50- 70%EtOAc/ Hex, obtain 4- ((6- aminopyridine -2- bases) epoxide) -2- methyl butyl- 2- alcohol (1.5g, 7.51mmol, 38.6% yield), it is pale solid, LCMS (m/z):197.29[M+H]⁺。

Synthesize 3- ((6- Aminopyrazine -2- bases) epoxide) -2,2- dimethyl propylene -1- alcohol

Under a nitrogen in the suspension of 0 DEG C of stirring to NaH (2.316g, 57.9mmol) in 1,4- dioxanes (20mL) Solution of middle dropwise addition 2,2- dimethyl propylenes -1, the 3- glycol (4.02g, 38.6mmol) in the dioxane of Isosorbide-5-Nitrae-(20mL), continues 10min.After 10min, solution of the 6- chloropyrazine -2- amine (5g, 38.6mmol) in the dioxane of Isosorbide-5-Nitrae-(20mL) is added dropwise at 0 DEG C, Continue 10min.Reactant mixture is heated into 48hr at 120 DEG C.TLC indicates a small amount of initiation material and product.Reaction is mixed Compound is poured into icy water (60mL), and water layer is extracted (2 × 100mL) with EtOAc.Organic layer is with salt water washing (50mL).Have Machine layer is through Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain crude product.Purified on column chromatography is purified, using 100-200 silica gel, (0-50%EtOAc/ petroleum ethers) obtains 3- ((6- Aminopyrazine -2- bases) epoxide) -2,2- dimethyl propylenes -1- as eluant, eluent Alcohol (1g, 4.95mmol, 12.84% yield), LCMS (m/z):198.00[M+H]⁺。

Synthesize 3- ((6- Aminopyrazine -2- bases) epoxide) propyl- 1- alcohol

Propyl- 1,3- is added in 0 DEG C of suspension to NaH (1.389g, 34.7mmol) in NMP (2mL) under a nitrogen Solution of the glycol (2.64g, 34.7mmol) in NMP (2mL), 1h is stirred at room temperature by reactant mixture.Then lasted at 0 DEG C Solution of the 6- chloropyrazine -2- amine (3g, 23.16mmol) in NMP (6mL) is added dropwise in 15min, and heats 16hr at 140 DEG C.Will be anti- Answer mixture to be cooled to room temperature, (50mL) is quenched with water and is extracted (3x150mL) with EtOAc.The organic matter of merging is washed with salt Wash, use anhydrous Na₂SO₄It is dried, filtered and concentrated, obtains crude product.Crude product is added into silicagel column and with (70%) EtOAc/ stones Oily ether elution.The fraction of collection is evaporated, acquisition compound 3- ((6- Aminopyrazine -2- bases) epoxide) propyl- 1- alcohol (1.5g, 8.21mmol, 35.5% yield), it is pale solid, LCMS (m/z):170.09[M+H]⁺。

Synthesize 6- (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethyoxyl) pyrazine -2- amine

NaH (60%) (0.556g, 23.16mmol) is added into 2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethanol at 0 DEG C In the solution of the stirring of (3.39g, 23.16mmol) in the dioxane of Isosorbide-5-Nitrae-(100mL), 30min is then stirred at room temperature and 0 DEG C add 6- chloropyrazine -2- amine (3g, 23.16mmol), then hold it in 80 DEG C of 16h.Reactant mixture is cooled to room Temperature, and be quenched with icy water (50mL), then distribute between icy water (20mL × 2) and ethyl acetate (20mL × 2). Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into crude compound, then it used to post color Spectrum purifying (uses 100-200 silicagel columns, eluted with 60% ethyl acetate/hexane), obtains 6- (2- ((tetrahydrochysene -2H- pyrans -2- Base) epoxide) ethyoxyl) pyrazine -2- amine (5g, 20.06mmol, 87% yield) is gluey grease, LCMS (m/z):240.13 [M+H]⁺。

Synthesize 4- ((6- Aminopyrazine -2- bases) epoxide) -2- methyl butyl- 2- alcohol

Under a nitrogen in the mixed of 0 DEG C of stirring to NaH (0.463g, 11.58mmol) in 1,4- dioxanes (5.00mL) 3- methyl butyl- 1, solution of the 3- glycol (1.206g, 11.58mmol) in the dioxane of Isosorbide-5-Nitrae-(5.00mL), 0 are added dropwise in suspension DEG C continue 10min.After 10min, 10min is lasted at 0 DEG C 6- chloropyrazine -2- amine (1.0g, 7.72mmol) is added dropwise in Isosorbide-5-Nitrae-Er Evil Solution in alkane (10.00mL).Reactant mixture is stirred into 16hr at 100 DEG C.Reaction process is monitored through TLC.By reaction mixing Thing pours into frozen water and extracted (3X25ml) with EtOAc, and organic solvent is through Na₂SO₄Dry and be concentrated in vacuo, obtain thick material.Thick thing Matter is purified through column chromatography, using silica gel (100-200 mesh), uses 50-70%EtOAc/ Hex, obtains 4- ((6- amino pyrroles Piperazine -2- bases) epoxide) -2- methyl butyl- 2- alcohol (1.0g, 5.04mmol, 65.3% yield), it is brown solid, LCMS (m/z): 199.08[M+H]⁺。

Synthesize (S) -3- bromo- 5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine

Cesium carbonate (37.0g, 114mmol) is absorbed in many neck RB.Then flask is cooled to 0 DEG C, and lasts 3 minutes Time be slowly added to METHYLPYRROLIDONE (NMP) (100mL).Gained reactant mixture is stirred under a nitrogen 15min.Then (S)-(2,2- dimethyl -1,3- dioxolane -4- bases) methanol is added dropwise at 0 DEG C in the time for lasting 5 minutes (10g,76mmol).1h is stirred at room temperature in the suspension.Add the bromo- 5- fluorine pyridines (7.62mL, 73.9mmol) of 3- suspension afterwards Become yellow solution.Resulting solution stirs 24hr at 75 DEG C.Reaction process is monitored through TLC 40%EtOAc/ hexanes.TLC refers to Show that initial substance exhausts, and new spot is formed after 24h.Reactive material is cooled to room temperature, is diluted with water (500mL).By water Layer is extracted with ethyl acetate (2X300mL).Organic layer is with salt water washing (250mL), through Na₂SO₄Dry and filter, be concentrated under reduced pressure, Obtain brown oil.Purified on column chromatography is purified, with the silica gel of 100-200 mesh sizes.By ladder of the post with EtOAc/ hexanes Degree elution.Desired compound 20%EtOAc/ Hex.Fraction containing pure compound is concentrated under reduced pressure, obtain (S)- (10g, 34.0mmol, 44.9% are received the bromo- 5- of 3- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine Rate), it is faint yellow sticky oil thing, LCMS (m/z):289.99[M+H]⁺。

Synthesize (R) -5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -3- amine

By the bromo- 5- of (R) -3- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine (50g, 174mmol), liquid ammonia (25mL, 1155mmol) is mounted in seal pipe.Then 0 DEG C add copper sulphate (II) (5.54g, 34.7mmol).Gained blue solution is heated to 120 DEG C, keeps 2hr.Reaction process is monitored through TLC10%MeOH/DCM, and TLC refers to Show that to form initial substance after new spot and 24h exhausts.After the completion of, reactive material is cooled to room temperature.It will be reacted with 20%NaOH The pH of material is changed into 10, uses NaCl saturations, is extracted with ethyl acetate (30mL*2).The organic layer of merging salt water washing (20mL), through Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain rough brown solid, it is used to triturated under ether and stirred 4 hours, Then filter, obtain (R) -5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -3- amine (35.4g, 146mmol, 84% yield), it is Light brown solid, LCMS (m/z):225.29[M+H]⁺。

Synthesize (S) -5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -3- amine

By the bromo- 5- of (S) -3- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine (10g, 34.7mmol), liquid ammonia (100mL, 4621mmol) is mounted in seal pipe.Gained brown solution is heated to 120 DEG C, holding 24hr.Reaction process is monitored through TLC (10%MeOH/DCM), and TLC indicates to be formed initial substance after new spot and 24h and exhausted. After the completion of, reactive material is cooled to room temperature.The pH of reactive material is changed into 10 with 20%NaOH, NaCl saturations is used, uses acetic acid second Ester extracts (30mL*2).The organic layer of merging is with salt water washing (20mL), through Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain (S) -5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -3- amine (6g, 25.8mmol, 74.2% Yield), it is Light brown solid, LCMS (m/z):225.10[M+H]⁺。

Synthesize (R) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -4- amine

Under a nitrogen in 0 DEG C of suspension to NaH (11.35g, 473mmol) in THF (100mL) be added dropwise (R)-(2, 2- dimethyl -1,3- dioxolane -4- bases) solution of the methanol (25g, 189mmol) in THF (150mL).Gained is mixed 1h is stirred at room temperature in suspension.In room temperature, 6- chlorine pyrimidine -4- amine (19.61g, 151mmol) is added dropwise to reactant mixture and by institute Obtain suspension and be heated to 90 DEG C, keep 48hr.(monitored after the completion of reaction through TLC, it shows a small amount of initial substance and observed New spot is formed in polar sites), reactant mixture is poured into frozen water (500mL), water layer extracted with EtOAc (2 × 1000mL).The organic matter of merging is through Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain light tan solid (rough thing).Thick material Purified (100-200,3%MeOH/DCM) through silicagel column.Fraction containing pure compound is merged and concentrated, desired production is obtained Thing (R) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -4- amine (13g, 53.9mmol, 28.5% yield), it is pale solid, also obtains impure compound (10g) .LCMS (m/z):226.17(M+H)⁺。

Synthesize (S) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -4- amine

Under a nitrogen in 0 DEG C of suspension to NaH (9.08g, 378mmol) in THF (150mL) be added dropwise (S)-(2, 2- dimethyl -1,3- dioxolane -4- bases) solution of the methanol (20g, 151mmol) in THF (200mL), gained is mixed 1h is stirred at room temperature in suspension.6- chlorine pyrimidine -4- amine (15.68g, 121mmol) is added dropwise to reactive material and by gained in room temperature Suspension is heated to 90 DEG C, keeps 48hr.(monitored after the completion of reaction through TLC, initiation material is completely depleted and observes new spot Point is formed in polar sites), reactive material is poured into frozen water (200mL) and is extracted with ethyl acetate (2X400mL).Merge Organic matter is through Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain light tan solid.The solid of acquisition is stirred in ether (200ml) Mix 30min.Filter and be dried in vacuo, obtain (S) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) phonetic Pyridine -4- amine (13g, 57.3mmol, 37.9% yield), it is light tan solid, LCMS (m/z):225.96[M+H]⁺。

Synthesize 2- ((tetrahydrochysene -2H- pyrans -4- bases) epoxide) pyrimidine -4- amine

27 DEG C to the tetrahydrochysene -2H- pyrans -4- alcohol (25g, 245mmol) under nitrogen gas stirring at tetrahydrofuran (THF) NaH (22.52g, 563mmol) is added in solution in (500mL), is lasted after 10min, 1hr in 27 DEG C of addition 2- chlorine pyrimidines -4- Amine (22.20g, 171mmol).Reactant mixture is stirred into 36hr at 85 DEG C.Reaction process is monitored by TLC.TLC indicates polarity Spot and initial substance.Reactive material is poured into 200ml icy waters, extracted with EtOAc (3X200ml), merging has Machine layer is through Na₂SO₄Dry, filter and be concentrated under reduced pressure, purified with column chromatography, use (60-120) mesh silica gel, initial substance uses 50% EtOAc/ Hex, required compound is eluted in 90%EtOAc/ hexanes, is concentrated the compound fraction merged, is obtained 2- ((tetrahydrochysene -2H- pyrans -4- bases) epoxide) pyrimidine -4- amine (9g, 39.9mmol, 16.31% yield), LCMS (m/z):196.00 [M+H]⁺。

Synthesize 4- (1- methyl isophthalic acid H- pyrazoles -4- bases) pyrimidine -2- amine

By 4- chlorine pyrimidine -2- amine (800mg, 6.18mmol), solid 1- methyl -4- (4,4,5,5- tetramethyls -1,3,2- bis- The amyl- 2- yls of oxa- boron heterocycle) -1H- pyrazoles (1927mg, 9.26mmol) and K₃PO₄(3932mg, 18.53mmol) is in 1,4- bis- Evil The solution of degassing is stirred at room temperature in alkane (20mL), water (5.00mL).Then PdCl is added₂(dppf)-CH₂Cl₂Adduct (756mg, 0.926mmol) and the 5min that deaerates.Then reactant mixture is stirred into 15h 30min at 80 DEG C.It is anti-by TLC monitorings Should.Reactant mixture is cooled to RT.Organic phase is evaporated, and is added 50mL water and is extracted with ethyl acetate (3x70ml), uses 50mL Saturated brine is washed, through Na₂SO₄Dry and be evaporated in vacuo, obtain crude product.Crude compound is (neutral through flash column chromatography Aluminum oxide, eluant, eluent：90%EtOAc/ petroleum ethers), obtain 4- (1- methyl isophthalic acid H- pyrazoles -4- bases) pyrimidine -2- amine (600mg, 3.42mmol, 55.5% yield), it is pale solid, LCMS (m/z):176.1[M+H]⁺。

Synthesize 4- (2- Jia Ji oxazole -5- bases) pyrimidine -2- amine

Mixed in room temperature to 4- chlorine pyrimidine -2- amine (1.5g, 11.58mmol) in 1,4- dioxanes (20mL) and water (5mL) In the solution of stirring in thing add potassium phosphate (3.69g, 17.37mmol) and 2- methyl -5- (4,4,5,5- tetramethyl -1,3, 2- dioxaborolan -2- base) oxazoles (2.421g, 11.58mmol).By reactive material nitrogen degassing 15min, add PdCl₂(dppf)-CH₂Cl₂Adduct (0.095g, 0.116mmol) simultaneously stirs gained reactant matter at 80 DEG C under a nitrogen 16hr, reaction process is monitored by TLC, and TLC indicates that to form multiple polarity spots and initial substance is depleted.Reactive material is dense Contracting, is then diluted with 50ml water and 60ml DCM, by Hi-flow, is separated organic layer, is extracted with DCM (2X50ml), merges Organic layer, through Na₂SO₄It is dried, filtered and concentrated, obtains crude compound.Crude product is used through combiflash chromatogram purifications Silicagel column (24g, 60%EtOAc/ petroleum ether).By gradient elution of the post with EtOAc/ hexanes.Desired compound uses 60% EtOAc/ Hex.Fraction containing pure compound is concentrated under reduced pressure, 4- (2- Jia Ji oxazole -5- bases) pyrimidine -2- amine is obtained (1.2g, 6.66mmol, 57.5% yield), it is pale solid.LCMS(m/z):177.11[M+H]⁺。

Synthesize (R) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- amine

Under a nitrogen 0 DEG C to NaH (11.67g, 292mmol) in METHYLPYRROLIDONE (NMP) (100mL) (R)-(2,2- dimethyl -1,3- dioxolane -4- bases) methanol (25.7g, 194mmol) is added dropwise in the suspension of stirring to exist Solution in METHYLPYRROLIDONE (NMP) (100mL), 10min is continued at 0 DEG C.After 10min, 6- chlorine pyrroles are added dropwise at 0 DEG C Solution of the pyridine -2- amine (25g, 194mmol) in METHYLPYRROLIDONE (NMP) (100mL), continues 10min.Will reaction Mixture heats 36hr at 100 DEG C.TLC indicates a small amount of initiation material and product.

Reactant mixture is poured into icy water (600mL), water layer is extracted (2 × 500mL) with EtOAc.Organic layer is used Water (3 × 300mL) washs to remove excessive NMP.Organic layer is through Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain crude product.Slightly Product is purified through column chromatography, using 100-200 silica gel, and (12-15%EtOAc/ petroleum ethers) obtains (R) -6- as eluant, eluent (10g, 44.6mmol, 22.93% are received ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- amine Rate), it is the liquid of yellow stiff.

Synthesize (S) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- amine

Under a nitrogen 0 DEG C to NaH (62.2g, 1556mmol) in METHYLPYRROLIDONE (NMP) (800mL) (S)-(2,2- dimethyl -1,3- dioxolane -4- bases) methanol (206g, 1556mmol) is added dropwise in the suspension of stirring to exist Solution in METHYLPYRROLIDONE (NMP) (300mL), continues 2h.Be stirred for after 10min, be added dropwise at 0 DEG C 6- chloropyridines- Solution of the 2- amine (200g, 1556mmol) in METHYLPYRROLIDONE (NMP) (300mL), continues 30min.Reaction is mixed Compound stirs 48hr at 120 DEG C.TLC indicates that initiation material exhausts.Reactant mixture is poured into icy water (2000mL), water Layer is extracted (3 × 1000mL) with EtOAc.The organic layer of merging is washed to remove excessive NMP with water (3 × 1000mL).Organic layer Through Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain crude product.Purified on column chromatography is purified, and uses (the elution of 100-200 silica gel Agent 12-15%EtOAc/ petroleum ethers), obtain desired pure product (S) -6- ((2,2- dimethyl -1,3- dioxolane - 4- yls) methoxyl group) pyridine -2- amine (75g, 325mmol, 20.92% yield), it is the liquid of clear yellow viscous.LCMS(m/z): 225[M+H]⁺。

Synthesize (R) -4- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -2- amine

Under a nitrogen in 0 DEG C of suspension to NaH (9.08g, 378mmol) in THF (150mL) be added dropwise (R)-(2, 2- dimethyl -1,3- dioxolane -4- bases) solution of the methanol (20g, 151mmol) in THF (250mL).Gained is suspended 1h is stirred at room temperature in liquid.4- chlorine pyrimidine -2- amine (15.68g, 121mmol) is added dropwise to reactant mixture and by gained in room temperature Suspension is heated to 90 DEG C, keeps 48hr.(monitored after the completion of reaction through TLC, initiation material exhausts and observes that new spot exists Polar sites are formed), reactant mixture is poured into frozen water (250mL), water layer is extracted (2 × 300mL) with EtOAc.Merge Organic matter is through Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain weak yellow liquid (rough thing).The thick material of acquisition is pure by post Change (100-200 silica gel), using 0-50%EtOAc- petroleum ethers, obtain (R) -4- ((2,2- dimethyl -1,3- dioxanes penta Alkane -4- bases) methoxyl group) pyrimidine -2- amine (13g, 57.0mmol, 37.7% yield), it is faint yellow solid, LCMS (m/z): 226.20[M+H]⁺。

Synthesize (S) -4- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -2- amine

It is added dropwise under a nitrogen in 0 DEG C of suspension to NaH (8.25g, 189mmol) in 1,4- dioxanes (200mL) (S)-(2,2- dimethyl -1,3- dioxolane -4- bases) methanol (10g, 76mmol) is in 1,4- dioxanes (50mL) Solution.1h is stirred at room temperature in gained suspension.4- chlorine pyrimidine -2- amine (7.84g, 60.5mmol) is added dropwise into reaction in room temperature to mix Gained suspension is simultaneously heated to 90 DEG C by compound, keeps 48hr.Reactant mixture is cooled to 28 DEG C, then distributed in water Between (200mL) and EtOAc (200mL).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and obtain filtrate evaporation Thick material (TLC eluant, eluents：Net ethyl acetate R_f0.3；UV activation).Crude compound purifies (100-200 mesh silicon through column chromatography Glue, is eluted in 60% ethyl acetate/hexane), obtain (S) -4- ((2,2- dimethyl -1,3- dioxolane -4- bases) first Epoxide) pyrimidine -2- amine (8.0g, 35.4mmol, 46.8% yield), it is faint yellow solid LCMS (m/z) 226.30 (M+H)⁺。

Synthesize (R)-(4- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -2-base) amino first Sour phenylester

Under nitrogen gas stirring room temperature to phenyl chloroformate (2.71g, 17.31mmol) and pyridine (1.724mL, (R) -4- ((2,2- dimethyl -1,3- dioxanes 21.31mmol) are added in the solution in dichloromethane (DCM) (50mL) Pentane -4- bases) methoxyl group) pyrimidine -2- amine (3.0g, 13.32mmol).Reactant mixture is stirred into 2hr at 28 DEG C.Pass through TLC Monitoring reaction.Reactant mixture is diluted with water into (75mL) to be extracted with DCM (2 × 75mL).Separation organic layer simultaneously uses Na₂SO₄Dry Dehydration, filters and high vacuum concentration, obtains crude product.Ether and pentane (3 are added into crude product:1) mixture and stirring 10min is simultaneously filtered, obtain compound (R)-(4- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine - 2- yls) phenyl carbamates (2.5g, 2.375mmol, 17.83% yield), LCMS (m/z):346.21[M+H]⁺。

Synthesize 2- (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethyoxyl) pyrimidine -4- amine

0 DEG C by NaH (0.741g, 30.9mmol) add to 2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethanol (4.51g, 30.9mmol) in the solution of the stirring in the dioxane of Isosorbide-5-Nitrae-(120mL), 30min is then stirred at room temperature and 2- is added at 0 DEG C Chlorine pyrimidine -4- amine (4g, 30.9mmol), then keeps 16h by it at 80 DEG C.Reactant mixture is cooled to room temperature, and uses ice Cold water (50mL) is quenched, and then distributes between icy water (50mL × 2) and ethyl acetate (50mL × 2).Separation is organic Layer, then through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into crude compound, then it is purified with column chromatography (makes 100-200 silica gel is used, post is eluted with 60% ethyl acetate/hexane), obtain 2- (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) Ethyoxyl) pyrimidine -4- amine (4g, 16.05mmol, 52.0% yield), it is glue grease LCMS (m/z):239.9[M+H ]⁺。

Synthesize 3- ((4- aminopyrimidine -2- bases) epoxide) -2,2- dimethyl propylene -1- alcohol

Under a nitrogen in the mixed of 0 DEG C of stirring to NaH (2.316g, 57.9mmol) in tetrahydrofuran (THF) (20mL) Solution of 2,2- dimethyl propylenes -1, the 3- glycol (4.02g, 38.6mmol) in tetrahydrofuran (THF) (20mL) is added dropwise in suspension, Continue 10min.After 10min, 2- chlorine pyrimidine -4- amine (5g, 38.6mmol) is added dropwise at 0 DEG C in tetrahydrofuran (THF) (20mL) Solution, continue 10min.Reactant mixture is heated into 16hr at 120 DEG C.TLC indicates a small amount of initiation material and product.Will Reactant mixture is poured into icy water (60mL), and water layer is extracted (2 × 100mL) with EtOAc.Organic layer salt water washing (50mL).Organic layer is through Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain crude product.Purified on column chromatography is purified, and is used 100-200 silica gel, eluant, eluent (0-50%EtOAc/ petroleum ethers) obtains 3- ((4- aminopyrimidine -2- bases) epoxide) -2,2- diformazans Base propyl- 1- alcohol (800mg, 4.04mmol, 10.47% yield), LCMS (m/z):198.09[M+H]⁺。

Under a nitrogen in the mixed of 0 DEG C of stirring to NaH (0.463g, 11.58mmol) in 1,4- dioxanes (5.00mL) 3- methyl butyl- 1, solution of the 3- glycol (1.206g, 11.58mmol) in the dioxane of Isosorbide-5-Nitrae-(5.00mL), 0 are added dropwise in suspension DEG C continue 10min.After 10min, 6- chloropyrazine -2- amine (1.0g, 7.72mmol) is added dropwise at 0 DEG C in the dioxane of Isosorbide-5-Nitrae - Solution in (10.00mL), continues 10min.Reactant mixture is stirred into 16hr at 100 DEG C.Reaction process is monitored by TLC. Reactant mixture is poured into frozen water and extracted (3X25ml) with EtOAc, organic solvent is through Na₂SO₄Dry and be concentrated in vacuo, obtain Thick material.Thick material is purified through column chromatography, using silica gel (100-200 mesh), is used 50-70%EtOAc/ Hex, is obtained 4- ((6- Aminopyrazine -2- bases) epoxide) -2- methyl butyl- 2- alcohol (1.0g, 5.04mmol, 65.3% yield), it is brown solid, LCMS(m/z):199.08[M+H]⁺。

Synthesize 4- ((4- aminopyrimidine -2- bases) epoxide) -2- methyl butyl- 2- alcohol

N35296-64

Under a nitrogen in the suspension of 0 DEG C of stirring to NaH (1.698g, 42.5mmol) in 1,4- dioxanes (100mL) 3- methyl butyl- 1 is added dropwise in liquid, solution of the 3- glycol (4.42g, 42.5mmol) in the dioxane of Isosorbide-5-Nitrae-(50mL) continues at 0 DEG C 15min.After 10min, 2- chlorine pyrimidine -4- amine (5.00g, 38.6mmol) is added dropwise at 0 DEG C, continues 15min.Reactant mixture is existed 100 DEG C of heating 16hr.Reaction process is monitored by TLC.Reactant mixture is poured into frozen water, concentrates, obtains as thick material Sticky material.Thick material is purified through column chromatography, using silica gel (60-120 mesh), is used 50-70%EtOAc/ Hex, is obtained To 4- ((4- aminopyrimidine -2- bases) epoxide) -2- methyl butyl- 2- alcohol (6.3g, 29.6mmol, 77% yield), it is canescence Solid, LCMS (m/z):198.30[M+H]⁺。

Synthesize (R) -5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- amine

N35281-46

To the chloro- 5- of (R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine in seal pipe In the solution of the stirring of (12g, 49.0mmol) in tetrahydrofuran (THF) (20mL) add ammonium hydroxide (300mL, 1926mmol) with copper sulphate (II) (1.566g, 9.81mmol).Reactant mixture is stirred into 18hr at 120 DEG C.Supervised by TLC Reaction process is surveyed, TLC is indicated to form polarity spot and be there is unreacted initial substance.Reactant mixture is diluted with water (300mL), is extracted (3x200mL) with EtOAc, and organic layer merges and uses water (100mL), saline solution (100mL) washing, organic Layer is through anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain (R) -5- ((2,2- dimethyl -1,3- dioxolane -4- Base) methoxyl group) pyrazine -2- amine (10g, 3.97mmol, 8.09% yield), it is the crude compound of yellow oily, LCMS (m/ z):226.13(M+H)⁺。

Synthesize (S) -2- chloro- 5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine

Under nitrogen gas stirring 0 DEG C to (S)-(2,2- dimethyl -1,3- dioxolane -4- bases) methanol (8.87g, Cesium carbonate (32.8g, 101mmol) 67.1mmol) is added in the suspension in DMF (DMF) (50mL), Gained reactant mixture is stirred into 1hr at 0 DEG C.2,5- dichloropyrazines (10g, 67.1mmol) are added thereto.Gained is reacted Mixture stirs 6hr at 100 DEG C.Reaction process is monitored by TLC.TLC indicates that initiation material exhausts, and forms new polarity spot Point, Rf is 0.3.Reactive material is cooled to room temperature, adds water (100mL) and is extracted with ethyl acetate (100mL).Organic layer water Wash (100mL × 2).Organic layer is through Na₂SO₄It is dried, filtered and concentrated, obtains thick material, it is light brown liquid.Crude product Add to silica gel (60-120) post and eluted with Hex/EtOAc.Fraction is collected, with 30%EtOAc/ Hex products.Concentration production Thing fraction, obtain the chloro- 5- of (S) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine (12g, 47.7mmol, 71.0% yield), it is light brown liquid, LCMS (m/z):244.90[M+H]⁺。

Synthesize (S) -5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- amine

In room temperature to the chloro- 5- of (S) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) under stirring at room temperature Methoxyl group) add in solution of the pyrazine (10g, 40.9mmol) in tetrahydrofuran (THF) (10mL) ammonium hydroxide (63.7mL, 409mmol) with copper sulphate (II) (3.26g, 20.44mmol).Reactant mixture is stirred 2 days in seal pipe at 130 DEG C.It is logical Cross TLC monitoring reaction process.TLC indicates that initiation material is depleted.Reactive material is cooled down to room temperature, is diluted with water (100mL), is used Ethyl acetate extracts (250mL × 2).Organic layer is through Na₂SO₄It is dried, filtered and concentrated, obtains crude compound, it is viscous for brown Thick compound.Crude product adds to silicagel column and eluted with DCM/EtOAc.The fraction of collection, is washed with 50%EtOAc/ petroleum ethers It is temporarily released from one's regular work thing.Enriched product fraction, obtain (S) -5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine - 2- amine (2g, 8.77mmol, 21.46% yield) (N35119-51-A2), it is light tan solid.NMR:In CDCl₃In, one Cause, LCMS (m/z):226.09[M+H]⁺。

Synthesize (S) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridazine -3- amine

Dripped under a nitrogen in 0 DEG C of suspension to KOtBu (12.99g, 116mmol) in 1,4- dioxanes (300mL) Plus the 20ml solution of (S)-(2,2- dimethyl -1,3- dioxolane -4- bases) methanol (4.08g, 30.9mmol).Gained is mixed 1h is stirred at room temperature in suspension.6- chlorine pyridazine -3- amine (5g, 38.6mmol) is added dropwise to reactant mixture in room temperature and gained is suspended Liquid is heated to 110 DEG C, keeps 16hr.(monitored after the completion of reaction through TLC, it shows a small amount of initial substance and observed newly Spot is formed in polar sites), reactant mixture is poured into frozen water (50mL), water layer is extracted (2 × 50mL) with EtOAc.Close And organic matter through Na₂SO₄Dry .LCMS (m/z):226.19[M+H]⁺。

Synthesize (R) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -3- amine

(R)-(2,2- dimethyl -1,3- dioxolane -4- bases) methanol (27.8g, 210mmol) is added at 0 DEG C In the solution of stirrings of the KOtBu (45.8g, 408mmol) in NMP (200mL), 1h is then stirred at room temperature, 0 DEG C is cooled to, 6- chloropyridine -3- amine (15g, 117mmol) is added, 110 DEG C are then heated to, 144h is kept.Reactant mixture is cooled to room Temperature is simultaneously distributed between water (500mL × 2) and EtOAc (200mL × 4).Organic layer is separated, then through anhydrous Na₂SO₄Dry, mistake Filter and filtrate evaporation is obtained into rough thing, and purified through column chromatography (using 100-200 silica gel, 50% ethyl acetate/hexane of post Elution), obtain (R) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -3- amine (8g, 35.1mmol, 30.1% yield), it is brown oil, LCMS (m/z):225.16[M+H]⁺。

Synthesize (S) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -3- amine

(S)-(2,2- dimethyl -1,3- dioxolane -4- bases) methanol (18.50g, 140mmol) is added at 0 DEG C In the solution of stirrings of the KOtBu (30.5g, 272mmol) in NMP (600mL), 1h is then stirred at room temperature, 0 DEG C is cooled to, 6- chloropyridine -3- amine (10.0g, 78mmol) is added, 110 DEG C are then heated to, 88h is kept.Reactant mixture is cooled to room Temperature is simultaneously distributed between water (50mL × 2) and EtOAc (100mL × 2).Organic layer is separated, then through anhydrous Na₂SO₄Dry, mistake Filter and filtrate evaporation is obtained into crude compound, it is jelly.(TLC:Eluant, eluent：100% ethyl acetate, R_f0.5；UV lives Change).Crude product is through flash column chromatography (silica gel：100-200 mesh), 50%EtOAc/ Hex is used, (S) -6- is obtained (10.0g, 41.7mmol, 53.6% are received ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -3- amine Rate), it is the material of dark viscous, LCMS (m/z) 225.0 (M+H)⁺。

Synthesize (R) -5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- amine

(R)-(2,2- dimethyl -1,3- dioxolane -4- bases) methanol (30.7g, 232mmol) is added at 0 DEG C In the solution of stirrings of the KOtBu (70.1g, 624mmol) in NMP (800mL), 1h is then stirred at room temperature and 0 DEG C is cooled to, Then 5- fluorine pyridine -2- amine (20g, 178mmol) is added, 110 DEG C are then heated to, 114h is kept.Reactant mixture is cooled down To room temperature and distribute between water (500mL × 2) and EtOAc (500mL × 4).Organic layer is separated, then through anhydrous Na₂SO₄It is dry It is dry, filter and filtrate evaporation is obtained into crude compound, then it is purified with column chromatography and (100-200 silica gel is used, with 80% Ethyl acetate/hexane elutes post), obtain (R) -5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrrole Pyridine -2- amine (10g, 40.1mmol, 22.50% yield), it is brown oil, LCMS:225.0(M+H).

Synthesize (S) -5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -2- amine

NaH (12.84g, 268mmol) is added into (S)-(2,2- dimethyl -1,3- dioxolane -4- bases) at 0 DEG C In the solution of stirring of the methanol (31.8g, 241mmol) in dimethyl sulfoxide (DMSO) (DMSO) (100mL), 1h is then stirred at room temperature And it is cooled to 0 DEG C, 5- fluorine pyridine -2- amine (15.0g, 134mmol) is added, 110 DEG C are then heated to, 60h is kept.Reaction is mixed Compound is cooled to room temperature, and distributes between water (50mL) and EtOAc (100mL).Organic layer is separated, then through anhydrous Na₂SO₄ Dry, filter and filtrate evaporation is obtained into crude compound (TLC:Eluant, eluent：100% ethyl acetate, R_f0.5；UV activation), Crude product is through flash column chromatography (silica gel：100-200 mesh) 50%EtOAc/ Hex is used, obtain (S) -5- ((2,2- bis- Methyl isophthalic acid, 3- dioxolane -4- bases) methoxyl group) pyridine -2- amine (7.2g, 32.1mmol, 23.99% yield), it is light Clear yellow viscous thing, LCMS (m/z):225.1(M+H)⁺。

Synthesize (R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -5- amine

Tetrahydrofuran (75mL) is added in NaH (5.56g, 232mmol) at 0 DEG C, at 0 DEG C, by (R)-(2,2- diformazans Base -1,3- dioxolane -4- bases) solution of the methanol (12.46mL, 100mmol) in tetrahydrofuran (50mL) adds to instead Answer mixture and reactant mixture is stirred into 1h at 28 DEG C.2- chlorine pyrimidine -5- amine (10g, 77mmol) is added in tetrahydrofuran (25mL) and stir 16hr at 70 DEG C.Reactant mixture is quenched with cold water (30mL) and extracted with ethyl acetate (3 × 80mL). Organic layer is washed with water (2 × 50mL) and saturated brine solution (50mL), through anhydrous Na₂SO₄It is dried, filtered and concentrated.Roughization Compound purifies the elution of (neutral alumina) product 40-45% ethyl acetate/hexanes through column chromatography, obtains (R) -2- ((2,2- bis- Methyl isophthalic acid, 3- dioxolane -4- bases) methoxyl group) pyrimidine -5- amine (6.5g, 28.3mmol, 36.6% yield), it is light Yellow solid, LCMS (m/z):226.0[M+H]⁺。

Synthesize (S) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -5- amine

In 0 DEG C of suspension to NaH (6.17g, 154mmol) in THF (100ml) add (S)-(2,2- dimethyl- 1,3- dioxolane -4- bases) solution of the methanol (13.26g, 100mmol) in THF (50ml), and by reactant mixture 1h is stirred at 25 DEG C.In the 0 DEG C of solution of 2- chlorine pyrimidine -5- amine (10g, 77mmol) in THF (50mL), and slowly adding thereto Heat to 80 DEG C and stirs 16hr at 80 DEG C.After the completion of reaction, reactant mixture is quenched with ammonium chloride (10mL) and uses ethyl acetate (3x20ml) is extracted.Separation organic layer simultaneously uses salt water washing, through Na₂SO₄Dry, filtered and be concentrated under reduced pressure, obtain thick thing Matter.The thick material triturated under ether, obtains (S) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) phonetic Pyridine -5- amine (5.0g, 19.77mmol, 25.6% yield), it is brown solid, LCMS (m/z):226.1[M+H]⁺。

Synthesize benzo [d] oxazole -2- aminocarbamic acid phenyl esters

In room temperature to pyridine (0.965mL, 11.93mmol) under nitrogen gas stirring, phenyl chloroformate (1.517g, Benzo [d] oxazole -2- amine (1g, 7.46mmol) 9.69mmol) is added in the solution in dichloromethane (DCM) (15mL).Will Reactant mixture stirs 3hr at 30 DEG C.Monitored and reacted by TLC.By reactant mixture with water (20mL), dichloromethane (2 × 50mL) dilute, separate organic layer.Organic layer washs (25mL) with saturated brine solution, through anhydrous Na₂SO₄Dry, filter and dense Contracting.Thick material is ground with 50% ether/hexane, and is analyzed, LCMS (m/z):255.19[M+H]⁺。

Synthesize (4- bromopyridine -2- bases) phenyl carbamates

Room temperature to phenyl chloroformate (4.98g, 31.8mmol) and Py (3.04mL, 37.6mmol) in DCM (40mL) Mixture in 4- bromopyridine -2- amine (5g, 28.9mmol) solution in DCM (30mL) is added dropwise.By resulting solution in room temperature Stir 2hr.(monitored after the completion of reaction through TLC, initiation material is completely depleted and observes new spot above initial substance just Point), saturated sodium bicarbonate solution (60mL) is added into reactive material and extracts water layer (2 × 70ML) with DCM.What is merged is organic Layer is through Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain pale solid.The solid of acquisition is stirred in petroleum ether (50mL), Filter and dry, obtain (4- bromopyridine -2- bases) phenyl carbamates (4g, 6.66mmol, 23.03% yield), it is white Color solid, LCMS (m/z):293.2(M+H)⁺。

Synthesize (2- amino -3,3,3- trifluoro propyls) t-butyl carbamate

Room temperature to be stirred at room temperature under a nitrogen 3,3,3- trifluoro propyl- 1,2- diamine dihydrochlorides (1g, 4.97mmol), Boc is added dropwise in solution of the DIPEA (2.61mL, 14.92mmol) in dichloromethane (DCM) (60mL)₂O The solution of (0.924mL, 3.98mmol) in dichloromethane (DCM) (10mL).3hr is stirred at room temperature in reactant mixture.It is logical Cross TLC monitoring reaction process.TLC indicates to form nonpolar spot, and initial substance is completely depleted.By reactant mixture cold water (50mL) dilutes and extracted (2 × 30ml) with DCM.The organic layer of merging is with salt water washing (10mL), through anhydrous Na₂SO₄Dry, Filter and be concentrated under reduced pressure, acquisition (2- amino -3,3,3- trifluoro propyls) t-butyl carbamate (800mg, 3.51mmol, 70.5% yield), it is colorless oil.

Synthesize (S) -2- chloro- 5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methyl) pyrazine

In the solution of 0 DEG C of stirring to cesium carbonate (492g, 1510mmol) in DMF (1000mL) add (S)-(2, 2- dimethyl -1,3- dioxolane -4- bases) methanol (133g, 1007mmol).Gained reactant mixture is stirred at room temperature 30min.Then solution of 2, the 5- dichloropyrazines (150g, 1007mmol) in DMF (500mL) is added at 0 DEG C, gained is reacted Mixture stirs 4h at 100 DEG C.(TLC systems:20% ethyl acetate/petroleum ether, R_f:0.5, UV activation).By reaction mixing Thing is diluted with icy water (500mL), is extracted with EtOAc (3 × 300mL).The organic layer of merging water (2 × 200mL) and salt The aqueous solution (100mL) is washed, through anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain crude compound.Crude compound is passed through Flash column chromatography (silica gel:100-200 mesh, eluant, eluent：10%EtOAc/ hexanes), obtain desired product (S) -2- chloro- 5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine (200g, 768mmol, 76% yield), it is Yellow liquid.LCMS(m/z):245.1[M+H]⁺。

To the chloro- 5- of (S) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine in seal pipe Ammonium hydroxide (1000mL, 6420mmol) and sulfuric acid are added in the solution of the stirring of (120g, 490mmol) in THF (30mL) Copper (II) (15.66g, 98mmol), then stirs 48h (TLC systems by gained reactant mixture at 120 DEG C:50% acetic acid second Ester/petroleum ether, Rf:0.4, UV activation).(300mL) is diluted with water in reactant mixture, is extracted with EtOAc (3x500mL). The organic layer of merging is washed with water (200mL) and saline solution (200mL), through anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure, Obtain crude compound.Thick material (uses 100-200 mesh silica gel and uses 40%EtOAc/ Hex through flash column chromatography Compound), obtain desired product (S) -5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine -2- Amine (65g, 280mmol, 57.2% yield), it is yellow crystalline solid.LCMS(m/z):226.13[M+H]⁺。

Synthesize (R) -2- chloro- 5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine

In the suspension of 0 DEG C of stirring to cesium carbonate (32.8g, 101mmol) in DMF (100mL) add (R)-(2, 2- dimethyl -1,3- dioxolane -4- bases) methanol (8.87g, 67.1mmol), 30min is then stirred at room temperature.Add Then 2,5- dichloropyrazines (10g, 67.1mmol), 4h is stirred by gained reactant mixture at 100 DEG C.(TLC systems:20% second Acetoacetic ester/hexane, Rf:0.5, UV activation).Reactant mixture is diluted with icy water (200mL), (3 are extracted with EtOAc ×100mL).The organic layer of merging is washed with water (2x50mL) and saline solution (50mL), through anhydrous Na₂SO₄Dry, filtering is simultaneously Be concentrated under reduced pressure, obtain the chloro- 5- of (R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine (12g, 43.8mmol, 65.3% yield), it is the compound of yellow oily.LCMS(m/z):244.99[M+H]⁺。

To the chloro- 5- of (R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine in seal pipe Ammonium hydroxide (400mL, 2568mmol) and sulphur are added in the solution of the stirring of (8g, 32.7mmol) in tetrahydrofuran (10mL) Reactant mixture is simultaneously stirred 48h by sour copper (II) (1.044g, 6.54mmol) at 120 DEG C.(TLC systems:50% ethyl acetate/ Hexane, Rf:0.4, UV activation).(200mL) is diluted with water in reactant mixture, is extracted with EtOAc (3x50mL).Merge Organic layer water (50mL), saline solution (50mL) washing, through anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain roughization Compound.Thick material through flash column chromatography (using 100-200 mesh silica gel and use 40%EtOAc/ Hex compound), receipts The pure fraction of collection is simultaneously concentrated under reduced pressure, and obtains (R) -5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrrole Piperazine -2- amine (2g, 8.65mmol, 26.4% yield), it is yellow crystalline solid.LCMS(m/z):226.10[M+H]⁺。

Synthesize (S) -2- chloro- 5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine

In the solution of 0 DEG C of stirring to 2- chlorine pyrimidine -5- alcohol (13g, 100mmol) in THF (100mL) add (S) - (2,2- dimethyl -1,3- dioxolane -4- bases) methanol (13.16g, 100mmol), triphenylphosphine (32.7g, 124mmol), DEAD (19.71mL, 124mmol) is subsequently added into, 4h is stirred at room temperature in reactant mixture.(TLC elution systems: 30%EtOAc/ petroleum ethers；R_f-0.5；UV activation).(50mL) is quenched with water in reactant mixture and is extracted into EtOAc (2x75mL).Separate organic layer and through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain crude product.By thick material through color Spectrum purifying (silica gel, eluant, eluent：20%EtOAc/ hexanes), obtain (S) -2- chloro- 5- ((2,2- dimethyl -1,3- dioxanes penta Alkane -4- bases) methoxyl group) pyrimidine (20g, 79mmol, 79% yield), it is pale solid.LCMS(m/z):245.10；[M+ H]⁺。

Synthesize (S) -5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -2- amine

By the chloro- 5- of (S) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine (10g, 40.9mmol) mixture with ammonia spirit (66.3ml, 1226mmol) heats 24h in seal pipe at 120 DEG C.(TLC is eluted System:100%EtOAc；R_f-0.2；UV activation).Reactant mixture is cooled to room temperature, (50mL) is quenched with water and extracts Into EtOAc (2x75mL).Organic layer is separated, through anhydrous sodium sulfate drying, filters and evaporates filtrate, obtain crude product, its For yellow solid.Crude compound is ground with pentane (50mL), obtains (S) -5- ((2,2- dimethyl -1,3- dioxanes penta Alkane -4- bases) methoxyl group) pyrimidine -2- amine (6.6g, 28.6mmol, 70.0% yield), it is pale solid.LCMS(m/z): 226.17；[M+H]⁺。

Synthesize (R) -2- chloro- 5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine

In the solution of 0 DEG C of stirring to 2- chlorine pyrimidine -5- alcohol (20g, 153mmol) in THF (100mL) add (R) - (2,2- dimethyl -1,3- dioxolane -4- bases) methanol (24.30g, 184mmol), triphenylphosphine (50.2g, 192mmol), it is subsequently added into DEAD (30.3mL, 192mmol) and 12h is stirred at room temperature in reactant mixture.(TLC elutions system System:70%EtOAc/ petroleum ethers；R_f-0.5；UV activation).(100mL) is quenched with water in reactant mixture and EtOAc is extracted into In (200mL).Separate organic layer and through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain crude product.Thick material is passed through Chromatogram purification (silica gel, eluant, eluent：35%EtOAc/ hexanes), obtain (R) -2- chloro- 5- ((2,2- dimethyl -1,3- dioxanes Pentane -4- bases) methoxyl group) pyrimidine (23g, 91mmol, 59.5% yield), it is white solid.LCMS(m/z):245.06；[M +H]⁺。

Synthesize (R) -5- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine -2- amine

By the chloro- 5- of (R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine (5g, 20.44mmol) mixture with ammonia spirit (50ml, 924mmol) heats 48h in seal pipe at 120 DEG C.(TLC elutions system System:100%EtOAc；R_f-0.2；UV activation).Reactant mixture is cooled to room temperature, (50mL) is quenched with water and is extracted into In DCM (2x75mL).Organic layer is separated, through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain (R) -5- ((2,2- bis- Methyl isophthalic acid, 3- dioxolane -4- bases) methoxyl group) pyrimidine -2- amine (2.7g, 11.5mmol, 57.5% yield), it is light Yellow solid.LCMS(m/z):226.02；[M+H]⁺。

Synthesize (S) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridazine -4- amine

Add in the suspension of 0 DEG C of stirring to potassium tert-butoxide (3.90g, 34.7mmol) in 1,4- dioxanes (50mL) Enter the mixture of (S)-(2,2- dimethyl -1,3- dioxolane -4- bases) methanol (2.75g, 20.84mmol), and will be anti- Mixture is answered to stir 1h at 25 DEG C in a nitrogen atmosphere.Then, 6- chlorine pyridazine -4- amine (1.5g, 11.58mmol) is added into reaction Mixture, 16h is stirred by gained reactant mixture at 110 DEG C.(TLC systems:Net ethyl acetate, Rf:0.3).By reaction mixing Thing is poured into icy water (40mL) and extracted (2x80mL) with EtOAc.The organic layer of merging with aqueous salt solu-tion (50mL), Through anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain crude compound.Thick material is through flash column chromatography (neutral alumina Aluminium, eluant, eluent：65% ethyl acetate/petroleum ether), obtain desired product (S) -6- ((2,2- dimethyl -1,3- dioxanes Pentane -4- bases) methoxyl group) pyridazine -4- amine (1.0g, 4.28mmol, 37.0% yield), it is white solid.LCMS(m/z): 226.20[M+H]⁺。

Synthesize (R) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridazine -4- amine

Add in the suspension of 0 DEG C of stirring to potassium tert-butoxide (7.80g, 69.5mmol) in 1,4- dioxanes (50mL) Enter (R)-(2,2- dimethyl -1,3- dioxolane -4- bases) methanol (5.20mL, 41.7mmol) and reactant mixture exists Under nitrogen atmosphere 1h is stirred at 25 DEG C.Then 6- chlorine pyridazine -4- amine (3g, 23.16mmol) is added into reactant mixture, by gained Reactant mixture stirs 16h at 110 DEG C.(TLC system ethyl acetate, Rf:0.3).Reactant mixture is poured into icy water (40mL) and extracted (2x80mL) with EtOAc.The organic layer of merging is with aqueous salt solu-tion (50mL), through anhydrous Na₂SO₄Dry, Filter and be concentrated under reduced pressure, obtain crude compound.Crude product is through flash column chromatography (neutral alumina, eluant, eluent：65% second Acetoacetic ester/petroleum ether), obtain desired product (R) -6- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridazine -4- amine (2.2g, 9.66mmol, 41.7% yield), it is pale solid.LCMS(m/z):226.05[M+H]⁺,Rt =1.00min.

The chloro- 7- of (4S) -8- (1- cyclopropyl -1H- pyrazoles -4- bases)-N- (2- (((R) -2,2- dimethyl -1,3- dioxas Pentamethylene -4- bases) methoxyl group) pyridin-4-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

To the chloro- 7- of (4S) -8- (1- cyclopropyl -1H- pyrazoles -4- bases) -2 being dissolved in tetrahydrofuran (THF) (30mL), 3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(400mg, 1.325mmol) is in nitrogen gas stirring Under 0 DEG C add triphosgene (393mg, 1.325mmol), TEA (0.924mL, 6.63mmol).Reactant mixture is stirred in room temperature Mix 30min.(R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- amine is added thereto (297mg, 1.325mmol) and stir 16h at 80 DEG C in seal pipe.So that reactant mixture reaches room temperature and uses 50ml water quenchings Go out and use 2x150ml ethyl acetate to extract, the organic layer of merging is through Na₂SO₄Dry and be concentrated under reduced pressure, obtain crude product.Crude product Through flash column chromatography (silica gel：100-200 mesh) and eluted with 2%MeOH-DCM, obtain (4S) -8- chloro- 7- (1- rings third Base -1H- pyrazoles -4- bases)-N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (210mg, 0.377mmol, 28.4% yield), LCMS (m/z):552.10[M+H]⁺。

Synthesize (4S) -8- chloro- N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -4- bases) -7- (2- methoxyl groups -5- (trifluoromethyl) pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza- 5 (2H)-formamides

At 25 DEG C to the chloro- 7- of (4S) -8- (2- methoxyl groups -5- (trifluoromethyl) pyridin-3-yl) -2,3,4,5- tetrahydrochysene -1, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(550mg, 1.483mmol) is in tetrahydrofuran (THF) (50mL) Stirring solution in add TEA (1.241mL, 8.90mmol) and triphosgene (440mg, 1.483mmol) and stir 1hr, plus Enter then (R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- amine (665mg, 2.97mmol), 15hr is heated at 65 DEG C.Reactant mixture is cooled to room temperature, is concentrated in vacuo and by residue distribution in water Between (20mL) and DCM (2X30mL).Separate organic layer and through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained rough Compound.Crude product is purified through GRACE, uses C-18 reversed-phase columns, mobile phase A:0.1% formic acid/water；B:ACN, product uses 81% ACN/0.1% formic acid/water elution.Evaporation solvent simultaneously uses saturation NaHCO₃Alkalization.Water layer is extracted with DCM.DCM layers through anhydrous Na₂SO₄Dry, filter and evaporate, obtain the pure chloro- N- of (4S) -8- (2- (((R) -2,2- dimethyl -1,3- dioxanes penta Alkane -4- bases) methoxyl group) pyridin-4-yl) -7- (2- methoxyl groups -5- (trifluoromethyl) pyridin-3-yl) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (400mg, 0.597mmol, 40.2% yield), its For pale solid, LCMS (m/z):621.22[M+H]⁺。

The chloro- 7- of (4S) -8- (2- methoxyl groups -5- (trifluoromethyl) pyridin-3-yl) -2,3,4,5- tetrahydrochysene -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza

In room temperature by chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazines of (4S) -7,8- two It is miscellaneous(1.0g, 4.35mmol), (2- methoxyl groups -5- (trifluoromethyl) pyridin-3-yl) boric acid (1.152g, 5.22mmol) and Cs₂CO₃The solution argon gas of the stirring of (4.25g, 13.04mmol) in 1,4- dioxanes (10mL) ＆ water (1mL) is de- in room temperature Gas 15 minutes.By PdCl₂(dppf)-CH₂Cl₂Adduct (0.355g, 0.435mmol) adds to reactant mixture.Then will reaction Mixture stirs 16hr at 110 DEG C.Monitored and reacted by TLC.Reactant mixture is cooled to room temperature, then filtered through diatomite And washed with EtOAc.Draw filtrate and concentrate and dissolved with EtOAc.EtOAc layers are successively used in combination with water and aqueous salt solu-tion Na₂SO₄Drying and dehydrating, filters and concentrates, obtain crude product.Crude product is purified through GRACE, uses C-18 reversed-phase columns, mobile phase A: 0.1% formic acid/water；B:ACN, product is eluted in 72%ACN/0.1% formic acid/water.Evaporation solvent simultaneously uses saturation NaHCO₃Alkali Change.Water layer is extracted with DCM.DCM layers through anhydrous Na₂SO₄Dry, filter and evaporate, obtain pure (4S) -8- chloro- 7- (2- first Epoxide -5- (trifluoromethyl) pyridin-3-yl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(650mg, 1.578mmol, 36.3% yield), it is brown solid, LCMS (m/z):371.45[M+H]⁺。

Synthesize (4S) -8- cyano group-N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -4- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides

25 DEG C to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diazaThe solution of the stirring of -8- formonitrile HCNs (300mg, 0.908mmol) in tetrahydrofuran (THF) (30mL) Middle addition TEA (0.760mL, 5.45mmol) and triphosgene (270mg, 0.908mmol) and stirring 1hr, then add (R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- amine (407mg, 1.816mmol), and at 65 DEG C Heat 15hr.Reactant mixture is cooled to room temperature, is concentrated in vacuo and by residue distribution in water (20mL) and DCM (2X30mL) Between.Separate organic layer and through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into crude compound.Crude product with it is another The same substance of batch merges, and is then purified through GRACE, uses C-18 reversed-phase columns, mobile phase A:0.1% formic acid/water；B:ACN, Product is eluted in 84%ACN/0.1% formic acid/water.Evaporation solvent simultaneously uses saturation NaHCO₃Alkalization.Water layer is extracted with DCM .DCM layer is through anhydrous Na₂SO₄Dry, filter and evaporate, obtain pure (4S) -8- cyano group-N- (2- (((R) -2,2- dimethyl -1, 3- dioxolane -4- bases) methoxyl group) pyridin-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- bridges Asia Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (250mg, 0.381mmol, 41.9% yield), it is Pale solid, LCMS (m/z):581.23[M+H]⁺。

Synthesize (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 8- formonitrile HCNs

In room temperature by the bromo- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza(500mg,1.301mmol)、Zn(CN)₂(764mg, 6.51mmol) and Zn (OAc)₂ The solution argon gas of the stirring of (287mg, 1.562mmol) in N,N-dimethylformamide (DMF) (20mL) deaerates in room temperature 15 minutes.By Pd₂(dba)₃(238mg, 0.260mmol) ＆DPPF (289mg, 0.521mmol) add to reactant mixture.Then will Reactant mixture stirs 16hr at 110 DEG C.Monitored and reacted by TLC.Reactant mixture is cooled to room temperature, then through diatomite Filter and washed with EtOAc.Draw filtrate and concentrate and dissolved with EtOAc.EtOAc layers of priority water and aqueous salt solu-tion are simultaneously Use Na₂SO₄Drying and dehydrating, filters and concentrates, obtain crude product.Crude product is purified through GRACE, uses C-18 reversed-phase columns, mobile phase A:0.1% formic acid/water；B:ACN, product is eluted in 60%ACN/0.1% formic acid/water.Evaporation solvent simultaneously uses saturation NaHCO₃ Alkalization.Water layer is extracted .DCM layers through anhydrous Na with DCM₂SO₄Dry, filter and evaporate, obtain pure (4S) -7- (3- (trifluoros Methyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 8- formonitrile HCNs (350mg, 1.016mmol, 78% yield), it is pale solid, LCMS (m/z):331.11[M+H]⁺。

Synthesis (4S) -8- bromo- 7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza

At 0 DEG C to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaBe added portionwise in the solution of the stirring of (1g, 3.28mmol) in chloroform (15mL) NBS (0.758g, 4.26mmol) and at 30 DEG C stir 5hr.Reactant mixture is concentrated in vacuo and by residue distribution in water (20mL) and DCM Between (2X20mL).Separate organic layer and through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain pure (4S) -8- bromo- 7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza (0.800g, 2.025mmol, 61.8% yield), it is pale solid, LCMS (m/z):384.07[M+H]⁺。

Synthesize (4S) -8- chloro- N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -4- bases) -7- (1- methyl isophthalic acid H- pyrazoles -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides

At 25 DEG C to the chloro- 7- of (4S) -8- (1- methyl isophthalic acid H- pyrazoles -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diazaAdded in the solution of (440mg, 1.596mmol) in tetrahydrofuran (THF) (10mL) TEA (1.112mL, 7.98mmol), is subsequently added into triphosgene (474mg, 1.596mmol), stir 1h and add (R) -2- ((2, 2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- amine (358mg, 1.596mmol), then add at 80 DEG C Hot 15h.Reactant mixture is cooled to 28 DEG C, then distributed between water (100mL) and EtOAc (2 × 100mL).Separation has Machine layer, then through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into crude product.Crude compound is purified through column chromatography, is made With neutral alumina, 1-2%MeOH/DCM obtains the chloro- N- of (4S) -8- (2- (((R) -2,2- dimethyl -1,3- as eluant, eluent Dioxolane -4- bases) methoxyl group) pyridin-4-yl) -7- (1- methyl isophthalic acid H- pyrazoles -4- bases) -3,4- dihydro -1,4- bridges Asia Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (330mg, 0.593mmol, 37.1% yield), LCMS (m/z):526.18[M+H]⁺。

Synthesize (4S) -8- chloro- 7- (1- methyl isophthalic acid H- pyrazoles -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza

In room temperature to chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazines of (4S) -7,8- two It is miscellaneous(4.300g, 18.69mmol) and 1- methyl -4- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) - 1H- pyrazoles (4.67g, 22.43mmol) adds Cs in the solution in 1,4- dioxanes (100.0mL) and water (25.00mL)₂CO₃ (18.27g, 56.1mmol) and the 20min that deaerates.Then by PdCl₂(dppf)-CH₂Cl₂Adduct (1.526g, 1.869mmol) Reactant mixture is added in room temperature and is deaerated 10 minutes again.Then reactant mixture is stirred to 110 DEG C, keeps 16h (TLC Eluant, eluent：10%MeOH/ ethyl acetate R_f:0.3；UV activation).Reactant mixture is cooled to room temperature, is diluted with water (100mL), is extracted with ethyl acetate (2X100mL).Organic layer is through anhydrous Na₂SO₄It is dried, filtered and concentrated, obtains crude product. Purified on column chromatography is purified, and using 100-200 mesh silica gel, 0-15% methanol/DCM is used as eluant, eluent.The fraction of collection is subtracted Pressure concentration, obtains the chloro- 7- of (4S) -8- (1- methyl isophthalic acid H- pyrazoles -4- bases) -2,3,4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-endo-methylene group pyrido [2,3-b] [1,4] diaza(1.5g, 3.41mmol, 18.27% yield).The purity of product is 62.76%, and it shows 34.97% initial substance.LCMS(m/z):276.53[M+H]⁺。

Synthesize (4S) -8- chloro- N- (2- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -4- bases) -7- (1- methyl isophthalic acid H- pyrazoles -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides

At 25 DEG C to the chloro- 7- of (4S) -8- (1- methyl isophthalic acid H- pyrazoles -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diazaAdded in the solution of (700mg, 2.54mmol) in tetrahydrofuran (THF) (100mL) TEA (1.769mL, 12.69mmol), is subsequently added into triphosgene (753mg, 2.54mmol), stir 1h and add (S) -2- ((2, 2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- amine (569mg, 2.54mmol), in 80 DEG C of heating 15h.Reactant mixture is cooled to 28 DEG C, then distributed between water (100mL) and EtOAc (2 × 100mL).Separation is organic Layer, then through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into crude product.Crude compound is purified through column chromatography, is used 100-200 mesh silica gel, and 2-6%MeOH/DCM is as eluant, eluent, obtains the chloro- N- of (4S) -8- (2- (((S) -2,2- diformazans Base -1,3- dioxolane -4- bases) methoxyl group) pyridin-4-yl) -7- (1- methyl isophthalic acid H- pyrazoles -4- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(350mg, 0.659mmol, 26.0% are received -5 (2H)-formamides Rate), it is light tan solid, LCMS (m/z):526.29[M+H]⁺。

To solid (4S) -7- (3- (difluoromethyl) phenyl) -8- methyl -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diazaSolid is added in the solution of (400mg, 1.327mmol) in tetrahydrofuran (THF) (5mL) Triphosgene (236mg, 0.796mmol), DIPEA (1.391mL, 7.96mmol) and it is stirred at room temperature under a nitrogen 30 minutes.With Afterwards thereto add (R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- amine (476mg, 2.124mmol), 15h is kept 30 minutes at 75 DEG C under the conditions of seal pipe.Pass through TLC and LCMS monitoring reactions.Reaction is mixed Compound is diluted with water (50ml) and is extracted with ethyl acetate (2 × 50ml).The organic layer anhydrous Na of merging₂SO₄Dry and depressurize Concentration, obtains crude product.Thick material is analyzed, LCMS (m/z):572.16[M+H]⁺。

The chloro- 7- of (4S) -8- (3- (difluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza

By chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas of (4S) -7,8- two (400mg, 1.738mmol), (3- (difluoromethyl) phenyl) boric acid (299mg, 1.738mmol) and potassium carbonate (721mg, 5.22mmol) stir and deaerated 15 points in room temperature with argon gas in the suspension in 1,4- dioxanes (10mL) ＆ water (1.5mL) Clock, by PdCl₂(dppf)-CH₂Cl₂Adduct (1420mg, 1.738mmol) adds to reactant mixture.Then by reactant mixture 16hr is stirred at 90 DEG C.Reactant mixture is cooled to room temperature, then filters and is washed (100mL) with EtOAc through diatomite.Inhale Take filtrate and concentrate and dissolved (50ml) with EtOAc.EtOAc layers successively washed with water (100ml) and saline solution (100mL) and Use Na₂SO₄Drying and dehydrating, filters and concentrates, obtain crude product.Purified on column chromatography is purified, using silica gel (100-200), with And eluted with 50%EtOAc/ hexanes (gradient system), obtain the chloro- 7- of desired product (4S) -8- (3- (difluoromethyl) benzene Base) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(500mg, 1.554mmol, 89% Yield), it is faint yellow solid, LCMS (m/z):302.11[M+H]⁺。

Synthesize (4S) -8- chloro- N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -4- bases) -7- (2- (trifluoromethyl) pyridin-4-yl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine- 5 (2H)-formamides

In room temperature to the chloro- 7- of (4S) -8- (2- (trifluoromethyl) pyridin-4-yl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diazaThe stirring of (350mg, 1.027mmol) in tetrahydrofuran (THF) (50mL) it is molten TEA (1.432mL, 10.27mmol) and triphosgene (610mg, 2.054mmol) are added in liquid.Reactant mixture is stirred in room temperature Mix 1h.Then by (R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- amine (230mg, 1.027mmol) add to reactant mixture.Gained reactant mixture is stirred into 16hr at 65 DEG C.Reaction process is monitored by TLC, It shows that initiation material is depleted.Reactant mixture is cooled to RT, (100mL) is diluted with water and (2* is extracted with ethyl acetate 60ml), the organic layer aqueous salt solu-tion of merging and through Na₂SO₄Dry, filter and be evaporated in vacuo, obtain crude compound. Crude compound is purified through column chromatography, using aluminum oxide, (2:8) eluted in EtOAc＆ hexanes.Fraction is concentrated in vacuo, obtained The chloro- N- of (4S) -8- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridin-4-yl) -7- (2- (trifluoromethyl) pyridin-4-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first Acid amides (250mg, 0.286mmol, 27.8% yield), it is light yellow gum thing, LCMS (m/z):591.2(M+H)⁺。

The chloro- 7- of (4S) -8- (2- (trifluoromethyl) pyridin-4-yl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza

At 0 DEG C to (4S) -7- (2- (trifluoromethyl) pyridin-4-yl) -2,3,4,5- tetrahydrochysene -1,4- bridges under nitrogen gas stirring Methylene pyridine simultaneously [2,3-b] [1,4] diazaIn the suspension of the stirring of (1.4g, 4.57mmol) in chloroform (20mL) Add NCS (0.610g, 4.57mmol).12h is stirred at room temperature in reactant mixture.Reactant mixture is diluted with water (100mL) and extracted (2*100ml) with EtOAc, the organic matter aqueous salt solu-tion of merging and through Na₂SO₄Dry, filtering is simultaneously It is evaporated in vacuo, obtains crude compound.Crude compound is purified through column chromatography, using neutral alumina, (1:9) EtOAc＆ oneself Eluted in alkane.Fraction is evaporated in vacuo, the chloro- 7- of (4S) -8- (2- (trifluoromethyl) pyridin-4-yl) -2 are obtained, 3,4,5- tetrahydrochysenes - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(700mg, 1.883mmol, 41.2% yield), it is gelatinization Compound, LCMS (m/z):340.97(M+H)⁺。

Synthesize (4S) -8- chloro- N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -4- bases) -7- (5- (trifluoromethyl) pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine- 5 (2H)-formamides

Under a nitrogen 0 DEG C in seal pipe to the chloro- 7- of (4S) -8- (5- (trifluoromethyl) pyridin-3-yl) -2,3,4,5- Tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(300mg, 0.880mmol) in THF (15mL) is stirred TEA (0.736mL, 5.28mmol) and triphosgene (261mg, 0.880mmol) are added in the solution mixed.By reactant mixture 25 DEG C stirring 1hr.Then at 0 DEG C by (R) -2- ((2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine -4- Amine (395mg, 1.761mmol) adds to reactive material, and reactant mixture is heated into 70 DEG C, and it is maintained into 16hr at 70 DEG C.Will Reactant mixture is cooled to room temperature, adds water (50mL) and is extracted (2x40mL) with EtOAc.The organic layer saline solution of merging Wash (50mL), organic layer is through Na₂SO₄Dry and concentrated, obtain crude compound.Crude compound is purified through column chromatography (C-18:With 65%ACN/1% ammonium bicarbonate aqueous solutions elute, obtain the chloro- N- of (4S) -8- (2- (((R) -2,2- dimethyl -1, 3- dioxolane -4- bases) methoxyl group) pyridin-4-yl) -7- (5- (trifluoromethyl) pyridin-3-yl) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (185mg, 0.313mmol, 35.6% yield), It is faint yellow solid, LCMS (m/z):591.20[M+H]⁺。

Synthesize (4S) -8- chloro- 7- (6- (trifluoromethyl) pyridine -2- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza

Under a nitrogen at 0 DEG C to (4S) -7- (6- (trifluoromethyl) pyridine -2- bases) -2,3,4,5- tetrahydrochysene -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diazaAdded in the solution of the stirring of (150mg, 0.490mmol) in chloroform (10mL) NCS(65.4mg,0.490mmol).Reactant mixture is reached room temperature and it is maintained into 16hr at 25 DEG C.Reactant mixture water (20mL) is quenched and is extracted (2x30mL) with EtOAc.The organic layer of merging aqueous salt solu-tion (50mL), organic layer warp Na₂SO₄Dry and concentrated, obtain crude compound.Crude product adds to silica gel (60-120) post and uses 50%EtOAC- oil Ether is eluted.The fraction of collection is concentrated, the chloro- 7- of (4S) -8- (6- (trifluoromethyl) pyridine -2- bases) -2 are obtained, 3,4,5- tetrahydrochysenes - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(60mg, 0.154mmol, 31.4% yield), it is faint yellow Solid, LCMS (m/z):341.15[M+H]⁺。

Synthesize 4- ((2S, 6R) -2,6- thebaine generations) pyridine -2- amine

Room temperature to 4- fluorine pyridine -2- amine (2.0g, 17.84mmol) (2S, 6R) -2,6- thebaines (4.11g, Potassium carbonate (7.40g, 53.5mmol), cuprous iodide (I) (0.340g, 1.784mmol) are added in suspension in 35.7mmol) And N, N'- dimethyl-ethylenediamine (0.315g, 3.57mmol).Gained reactant mixture is stirred into 1hr in microwave at 110 DEG C (TLC systems:10%MeOH/DCM, R_f-:0.1；UV activation).Reactant mixture is quenched into (20mL) in frozen water to be used in combination EtOAc extracts (3X40mL).The organic layer aqueous salt solu-tion of merging, through anhydrous Na₂SO₄Dry, filter and evaporate and obtain thick Produced compounds.Crude compound is purified through column chromatography, using neutral alumina, is eluted, is obtained in 50%EtOAc/ petroleum ethers 4- ((2S, 6R) -2,6- thebaine generations) pyridine -2- amine (1.0g, 4.72mmol, 26.4% yield), it is pale yellow colored solid Body, LCMS (m/z):208.05[M+H]⁺。

Synthesize (4S)-N- (4- chlorine pyrimidine -2-base) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In -78 DEG C of stirrings to 4- chlorine pyrimidine -2- amine (2.61g, 20.14mmol) in tetrahydrofuran (THF) (60mL) Solution in add LiHMDS (67.1mL, 67.1mmol) and -78 DEG C stir 1hr, then add (4S) -7- (2- methyl pyrroles Pyridine -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formic acid phenylesters (5g, 13.43mmol), then 18hr is stirred at 25 DEG C.By reactant mixture distribution between water (50mL) and EtOAc (800mL).Point From organic layer, then through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain crude product.By residue triturated under ether (3 ×30mL).Gained solid is filtered, (4S)-N- (4- chlorine pyrimidine -2-base) -7- (2- picoline -4- bases) -3,4- bis- is obtained Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(2.1g, 4.86mmol, 36.2% are received -5 (2H)-formamides Rate), it is faint yellow solid, LCMS (m/z):408.17[M+H]⁺。

(4S)-N- (4- (2- ((S) -1,4- dioxo spiros [4.5] decyl- 2- yls) ethyl) pyridine -2- bases) -7- (2- methyl Pyridin-4-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature in a nitrogen atmosphere to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diazaThe stirring of (0.962g, 3.81mmol) in tetrahydrofuran (THF) (40mL) Triphosgene (0.905g, 3.05mmol) and TEA (1.594mL, 11.44mmol) are added in solution.Gained reactant mixture is existed 1hr is stirred at room temperature.(S) -4- (2- (1,4- dioxo spiros [4.5] decyl- 2- yls) ethyl) pyridine -2- is added into reactant mixture Solution of the amine (1.0g, 3.81mmol) in tetrahydrofuran (THF) (20mL).By gained reactant mixture in 70 DEG C of stirrings 16hr.Reaction process is monitored by TLC.(50mL) is diluted with water in reactant mixture, is extracted with EtOAc (3 × 50mL).Organic layer Merge and use aqueous salt solu-tion (30mL), through anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain crude compound.Thick thing Matter purifies (100-200 mesh silica gel, eluant, eluent is 4%MeOH/DCM) through column chromatography, obtain (4S)-N- (4- (2- ((S)-Isosorbide-5-Nitrae- Dioxo spiro [4.5] decyl- 2- yls) ethyl) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (800mg, 1.219mmol, 32.0% yield), it is palm fibre Color solid, LCMS (m/z):541.27[M+H]⁺。

Synthesize (1H- imidazoles -1- bases) ((4S) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-yls) ketone

TEA (1.370mL, 9.83mmol) is added into (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- four at 28 DEG C Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(1g, 3.28mmol) is in tetrahydrofuran (THF) (40mL) Stirring solution in.Reactant mixture is stirred into 1h at 28 DEG C.CDI (1.062g, 6.55mmol) is added into reactant mixture, And stir 12h at 60 DEG C.Reactant mixture is cooled to 28 DEG C, then distributed between water (20mL) and EtOAc (2X35mL). EtOAc layers of separation, then through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain thick material.Thick material is purified, second is used Ether (2X25mL) is washed, and obtains pure (1H- imidazoles -1- bases) ((4S) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro-Isosorbide-5-Nitraes - Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-yls) ketone (850mg, 2.097mmol, 64.0% yield), It is pale solid.LCMS(m/z):400.30[M+H]⁺。

Synthesize (4S) -8- chloro- N- (5- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrroles Pyridine -3- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C by the chloro- 7- of (4S) -8- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza(0.5g, 1.744mmol), triethylamine (1.458mL, 10.46mmol) are absorbed in tetrahydrofuran (THF) in (20mL), gained yellow solution is stirred into 10min.Then 0 DEG C it is disposable add triphosgene (0.517g, 1.744mmol).45min is stirred at room temperature in gained yellow suspension.At 0 DEG C by (R) -5- ((2,2- dimethyl -1,3- dioxas Pentamethylene -4- bases) methoxyl group) THF (4mL) solution of pyridine -3- amine (0.391g, 1.744mmol) adds to above-mentioned yellow and is suspended Liquid, lasts 5min time.Gained yellow suspension is heated to 70 DEG C, keeps 24hr.Reaction process is through TLC10%MeOH/DCM Monitoring, TLC forms multiple spots after indicating 24h.Reactive material is cooled to room temperature, and (20mL), ethyl acetate is diluted with water (30mL*2).The organic layer of merging is with salt water washing (15mL), through Na₂SO₄Dry filter, is concentrated under reduced pressure, and obtains brown viscous oil Shape thing.Crude reaction material is subjected to next step, LCMS (m/z) without any purifying:537.00[M+H]⁺。

Synthesis 4- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) -1- (2,2,2- trifluoroethyls) - 1H- pyrazoles

Room temperature to 4- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) -1H- pyrazoles (5.0g, Cs2CO3 (16.79g, 51.5mmol) 25.8mmol) is added in the solution in DMF (DMF) (50mL), Then trifluoromethanesulfonic acid 2,2,2- trifluoroethyls ester (4.45mL, 30.9mmol) is added dropwise in room temperature.By reactant mixture at 100 DEG C Stir 3hr.(100mL) is diluted with water in reactant mixture and is extracted (2 × 100mL) with EtOAc, the organic matter cold water of merging (3 × 100ml) and saline solution (100mL) are washed, through Na₂SO₄Dry, filter and be evaporated in vacuo, obtain 4- (4,4,5,5- tetra- Methyl isophthalic acid, 3,2- dioxaborolan -2- bases) -1- (2,2,2- trifluoroethyls) -1H- pyrazoles (1.8g, 3.39mmol, 13.16% yield), it is canescence jelly liquid.

COMPOUNDS EXAMPLE

Embodiment 1

Synthesize (4S) -7- (6- picoline -3- bases)-N- (pyridin-3-yl) -8- (trifluoromethyl) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -7- (6- picoline -3- bases) -8- (trifluoromethyl) -2,3,4,5- tetrahydrochysene -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza(200mg, 0.624mmol), triphosgene (111mg, 0.375mmol) are in tetrahydrochysene furan Solid pyridine -3- amine (88mg, 0.937mmol) and DIPEA (242mg, 1.873mmol) are added in the solution muttered in (10mL), Then it is stirred for 30min.Reactant mixture is heated to 75 DEG C in seal pipe, 16h is kept.Room temperature is allowed to cool to, is fallen Enter saturation NaHCO₃In solution (100mL), and (3x50mL) is extracted with ethyl acetate.The organic layer of merging is through anhydrous Na₂SO₄It is dry It is dry, it is concentrated under reduced pressure, obtains crude compound, by it by flash column chromatography, obtain (4S) -7- (6- picolines -3- Base)-N- (pyridin-3-yl) -8- (trifluoromethyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (127mg, 45.3% yield), it is pale solid (TLC:Eluant, eluent, 5% methanol/ethyl acetate； R_f=0.3), LCMS (m/z):441.23[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 12.67 (s, 1H), 8.69 (d, J=2.19Hz, 2H), 8.37-8.16 (m, 4H), 8.03 (ddd, J=8.33,2.63,1.53Hz, 2H), 7.89 (s, 2H), 7.76 (dd, J=8.00,2.30Hz, 2H), 7.32 (d, J=7.89Hz, 2H), 7.27-7.20 (m, 2H), 5.70 (dd, J=5.81,3.18Hz, 2H), 3.38-3.12 (m, 4H), 3.11-3.02 (m, 2H), 2.68 (s, 6H), 2.37 (dddd, J=14.20,10.03,6.03,4.17Hz, 2H), 2.24-2.01(m,2H)。

Embodiment 2

Synthesize (4S) -8- bromo- 7- (6- picoline -3- bases)-N- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the bromo- 7- of (4S) -8- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diazaNaH is added in the solution of the stirring of (350mg, 1.057mmol) in THF (20mL) (38.0mg,1.585mmol).Then reactant mixture is stirred into 1h at 30 DEG C, adds 3- (pyridine -2- bases) -2H- pyridos [1,2-a] [1,3,5] triazine -2,4 (3H)-diketone (305mg, 1.268mmol).Reactant mixture is stirred into 15h at 70 DEG C.Make It is cooled to RT and reactant mixture is poured into cold water (20mL) and is extracted with ethyl acetate (2X20mL).The organic layer of merging Through anhydrous Na₂SO₄Dry and be concentrated in vacuo, obtain crude compound.Crude mixture is through flash column chromatography (silica gel：100- 200 mesh, eluant, eluent：3%MeOH/EtOAc), 500mg is obtained, it is 90% to determine purity by LCMS.Half pure compound is through system Standby type HPLC purifying (posts：XS PHENYL HEXYL (250X4.6mm, 5 μ), mobile phase:A:0.01M ammonium hydrogen carbonate B:ACN, ladder Degree：Time/%B:0/10,1/10,10/50,15/50,18/98,20/98,20.1/10,25/10, column temperature：Environment temperature, stream Speed:1.0ml/min, diluent：ACN), the bromo- 7- of (4S) -8- (6- picoline -3- bases)-N- (pyridine -2- bases) -3,4- are obtained Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamide (275mg, 0.61mmol, 47% Yield), it is pale solid (TLC:10%MeOH/ ethyl acetate, R_f:0.5), LCMS (m/z):451.13[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 12.89 (s, 1H), 8.88 (d, J=2.19Hz, 1H), 8.32 (d, J= 2.41Hz, 1H), 8.26 (dd, J=4.82,1.32Hz, 1H), 8.04 (d, J=8.20Hz1H), 7.83 (s, 1H), 7.66 (t, J =7.20Hz 1H), 7.33 (d, J=7.89Hz, 1H), 6.98 (m, 1H), 5.67 (dd, J=6.03,3.18Hz, 1H), 3.49 (d, J=5.04Hz, 1H), 3.37-3.17 (m, 2H), 3.02 (dd, J=12.06,3.29Hz, 1H), 2.67 (s, 3H), 2.34 (dddd, J=14.14,10.08,6.03,3.95Hz, 1H), 2.18-2.01 (m, 1H).

Embodiment 3

Synthesize (4S) -8- chloro- 7- (6- picoline -3- bases)-N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

By Nicotinicum Acidum (180mg, 1.465mmol), DPPA (537mg, 1.953mmol) and triethylamine (0.680mL, 4.88mmol) solution in THF (20mL) stirs 2h at 30 DEG C, adds the chloro- 7- of (4S) -8- (6- picoline -3- bases) -2, 3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(280mg,0.976mmol).By reaction mixing Thing stirs 14h at 70 DEG C.Reactant mixture is poured into cold water (50mL) and is extracted with ethyl acetate (2X30mL).What is merged has Machine layer is through anhydrous Na₂SO₄Dry and be concentrated under reduced pressure, obtain crude product.Crude mixture is through flash column chromatography (silica gel：100- 200 mesh, eluant, eluent：3%MeOH/EtOAc), obtain the chloro- 7- of (4S) -8- (6- picoline -3- bases)-N- (pyridin-3-yl) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (90mg, 0.22mmol, 15% yield), it is white solid (TLC:10%MeOH/ ethyl acetate, R_f:0.3), LCMS (m/z):407.25[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 12.63 (s, 1H), 8.90 (d, J=1.97Hz, 1H), 8.36 (d, J= 2.19Hz,1H),8.31-8.21(m,1H),8.21-8.02(m,1H),8.02–7.92(m,1H),7.66(s,1H),7.34(d, J=8.11Hz, 1H), 7.27-7.21 (m, 1H), 5.67 (dd, J=5.92,3.07Hz, 1H), 3.37-3.11 (m, 2H), 3.11–2.92(m,2H),2.67(s,3H),2.42-2.22(m,1H),2.21-2.04(m,1H)。

Embodiment 4

At 0 DEG C to (4S) -7- (6- picoline -3- bases) -8- (trifluoromethyl) -2,3,4,5- tetrahydrochysene -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diazaTriphosgene is added in the solution of (200mg, 0.624mmol) in THF (10mL) (111mg, 0.375mmol), is then stirred at room temperature 30min.Then add pyridine -3- amine (88mg, 0.937mmol) and DIPEA(242mg,1.873mmol).Reactant mixture is heated into 16h at 75 DEG C in seal pipe.Reactant mixture is poured into full And NaHCO₃In solution (100mL) and it is extracted with ethyl acetate (3X50mL).The organic layer of merging is through anhydrous Na₂SO₄Dry and subtract Pressure concentration, obtains half pure compound.It is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：3%MeOH/ DCM), (4S) -7- (6- picoline -3- bases)-N- (pyridin-3-yl) -8- (trifluoromethyl) -3,4- dihydros-Isosorbide-5-Nitrae-bridge is obtained Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (127mg, 0.288mmol, 45.3%), it is ash White solid (TLC:R_f=0.3,5% methanol/ethyl acetate), LCMS (m/z):441.23[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 12.67 (s, 1H), 8.69 (d, J=2.19Hz, 1H), 8.37-8.16 (m, 2H), 8.03 (ddd, J=8.33,2.63,1.53Hz, 1H), 7.89 (s, 1H), 7.76 (dd, J=8.00,2.30Hz, 1H), 7.32 (d, J=7.89Hz, 1H), 7.27-7.20 (m, 1H), 5.70 (dd, J=5.81,3.18Hz, 1H), 3.38-3.12 (m, 3H), 3.02-3.11 (m, 1H), 2.68 (s, 3H), 2.39 (s, 3H), 2.37 (dddd, J=14.20,10.03,6.03, 4.17Hz,1H),2.24-2.01(m,1H)。

Embodiment 5

Synthesize (4S) -8- fluoro- 7- (6- picoline -3- bases)-N- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Successively triethylamine (0.516mL, 3.70mmol) and triphosgene (220mg, 0.740mmol) are added in room temperature The fluoro- 7- of (4S) -8- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneousIn the solution of the stirring of (200mg, 0.740mmol) in tetrahydrofuran (THF) (15mL, in seal pipe) and stir 30min, then adds pyridine -2- amine (84mg, 0.888mmol), and 15h is heated at 80 DEG C.Reactant mixture is cooled to room temperature, Then distribute between water (25mL) and EtOAc (40mL), separate organic layer and through anhydrous Na₂SO₄Dry, filter and by filtrate Evaporation, obtains crude compound.Crude mixture purifies (mobile phase-A through preparation HPLC:10mM ammonium hydrogen carbonate (aqueous solution): Mobile phase-B：Acetonitrile.Post：(100x25) 5 μ of YMC fillings:Method-T/%B:0.1/50,9/50,9.1/100,12/100, 12.1/50. flow velocity：20ml/min. eluant, eluents (solubility)：THF+ACN+MeOH.), (4S) -8- fluoro- 7- (6- first is obtained Yl pyridines -3- bases)-N- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (65mg, 0.165mmol, 22.33% yield), it is pale solid (TLC eluant, eluents：Net ethyl acetate R_f:0.5), LCMS (m/z):391.23[M+H]⁺。

¹H NMR(400MHz,CDCl₃)δppm:13.09 (s, 1H), 9.18 (s, 1H), 8.47 (dd, J=8.4,2.41Hz, 1H), 8.36-8.34 (m, 1H), 8.14 (d, J=7.6Hz, 1H), 7.71-7.66 (m, 1H) 7.42 (d, J=10.4Hz, 1H), 7.35 (d, J=8Hz, 1H), 5.68 (dd, J=6.03,3.2Hz, 2.8Hz, 1H), 7.01-6.96 (m, 1H), 3.36-3.14 (m,3H),3.02-2.88(m,1H),2.66(s,3H),2.30-2.29(m,1H),2.10–2.04(m,1H)。

Embodiment 6

Synthesize (4S) -8- bromo- 7- (6- picoline -3- bases)-N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the bromo- 7- of (4S) -8- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diazaTriphosgene is added in the solution of the stirring of (250mg, 0.755mmol) in THF (10mL) (134mg,0.453mmol).Then by reactant mixture 30 DEG C stir 30min and add pyridine -3- amine (142mg, 1.510mmol).Reactant mixture is stirred into 15h at 70 DEG C.Allow to cool to RT and reactant mixture is poured into cold water (20mL) In and be extracted with ethyl acetate (2X20mL).The organic layer of merging is through anhydrous Na₂SO₄Dry and be concentrated in vacuo, obtain rough chemical combination Thing.Crude mixture is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：3%MeOH/EtOAc), 500mg is obtained, It is 90% to determine purity by LCMS.Half pure compound purifies (post through preparation HPLC：XS PHENYL HEXYL (250X4.6mm, 5 μ), mobile phase:A:0.01M ammonium hydrogen carbonate B:ACN, gradient：Time/%B:0/10,1/10,10/50,15/ 50,18/98,20/98,20.1/10,25/10, column temperature：Environment temperature, flow velocity:1.0ml/min, diluent：ACN), obtain The bromo- 7- of (4S) -8- (6- picoline -3- bases)-N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza- 5 (2H)-formamides (195mg, 0.413mmol, 54.7% yield), it is yellow solid (TLC: 10%MeOH/ ethyl acetate, R_f:0.4), LCMS (m/z):451.17[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 12.60 (s, 1H), 8.88 (d, J=2.19Hz, 1H), 8.36-8.22 (m, 2H), 8.12-8.02 (m, 1H), 7.90 (dd, J=8.00,2.30Hz, 1H), 7.83 (s, 1H), 7.33 (d, J= 7.89Hz, 1H), 7.22 (dd, J=8.44,4.71Hz, 1H), 5.67 (dd, J=6.03,3.18Hz, 1H), 3.39-3.09 (m, 3H), 3.02 (dd, J=12.06,3.29Hz, 1H), 2.67 (s, 3H), 2.34 (dddd, J=14.14,10.08,6.03, 3.95Hz,1H),2.18-2.04(m,1H)

Embodiment 7

Synthesize (4S) -8- fluoro- 7- (6- picoline -3- bases)-N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Triethylamine (0.516mL, 3.70mmol) and DPPA (407mg, 1.480mmol) are successively added into nicotinic acid in room temperature Added in the solution of the stirring of (109mg, 0.888mmol) in THF (10mL), after 2h (4S) -8- fluoro- 7- (6- picolines - 3- yls) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(200mg, 0.740mmol), so After be heated to 80 DEG C, keep 16h.Reactant mixture is cooled to room temperature and distributed between water (25mL) and EtOAc (30mL), Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and evaporate filtrate decompression, obtain crude compound.Crude mixture (mobile phase-A is purified through preparation HPLC:10mM ammonium hydrogen carbonate (aqueous solution):Mobile phase-B：Acetonitrile post：YMC fillings (100x25)5μ:Method-T/%B:0.1/40,9/40,9.1/100,13/100,13.1/40. flow velocity：20ml/min eluant, eluents： THF+ACN+MeOH), the fluoro- 7- of (4S) -8- (6- picoline -3- bases)-N- (pyridin-3-yl) -3,4- dihydros-Isosorbide-5-Nitrae-bridge is obtained Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (52mg, 0.133mmol, 17.93% yield), its For pale solid (TLC eluant, eluents：5%MeOH/DCM, Rf:0.2), LCMS (m/z):391.30[M+H]⁺。

¹H NMR(400MHz,CDCl₃-d):δ ppm 12.65 (s, 1H), 8.96 (s, 1H), 8.48 (d, J=2.4Hz, 1H), 8.28 (dd, J=4.6,1.2Hz, 1H), 8.18-8.14 (m, 1H), 8.05-7.89 (m, 1H), 7.44 (d, J=10Hz, 1H), 7.35 (d, J=8Hz, 1H), 7.28-7.22 (m, 1H), 5.68 (dd, J=6,3.2Hz, 1H), 3.36-3.14 (m, 3H), 3.05-3.00 (m, 1H), 2.66 (s, 3H), 2.39-2.31 (m, 1H), 2.13-2.04 (m, 1H).

Embodiment 8

Synthesize (4S) -8- methyl -7- (6- picoline -3- bases)-N- (pyridine -2- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -8- methyl -7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diazaTriphosgene is added in the solution of the stirring of (200mg, 0.751mmol) in THF (15ml) (134mg, 0.451mmol), is then stirred to room temperature, continues 1h.Be subsequently added pyridine -2- amine (106mg, 1.126mmol) and DIPEA(291mg,2.253mmol).Reactant mixture is heated into 16h at 75 DEG C in seal pipe.Reactant mixture is poured into full And NaHCO₃Solution (50mL) is simultaneously extracted with ethyl acetate (3x50mL).The organic layer of merging is through anhydrous Na₂SO₄Dry and depressurize Concentration, obtains crude compound.Crude compound is through flash column chromatography (silica gel：100-200 mesh, uses 1% methanol/bis- The gradient mixture of chloromethanes), obtain (4S) -8- methyl -7- (6- picoline -3- bases)-N- (pyridine -2- bases) -3,4- bis- Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamide (150mg, 0.386mmol, 51.4% Yield), it is white solid (TLC systems:R_f:0.3, eluant, eluent：5% ethanol/methylene), LCMS (m/z):387.29[M +H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.32 (s, 1H), 8.79 (d, J=2.41Hz, 1H), 8.27 (ddd, J =4.82,1.97,0.88Hz, 1H), 8.15-8.06 (m, 2H), 7.64 (td, J=7.78,1.97Hz, 1H), 7.44 (s, 1H), 7.33 (d, J=8.11Hz, 1H), 6.94 (ddd, J=7.29,4.88,0.99Hz, 1H), 5.65 (dd, J=5.92,3.07Hz, 1H), 3.37-3.12 (m, 3H), 2.99 (dd, J=12.06,3.29Hz, 1H), 2.65 (s, 3H), 2.41 (s, 3H), 2.35- 2.25(m,1H),2.16-2.06(m,1H)。

Embodiment 9

Synthesize (4S) -9- methyl -7- (6- picoline -3- bases)-N- (pyridine -2- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Sodium hydride (NaH) (0.270g, 6.76mmol) is added into (4S) -9- methyl -7- (6- methyl in room temperature under a nitrogen Pyridin-3-yl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(0.300g, 1.126mmol) in the solution of the stirring in tetrahydrofuran (30mL).Reactant mixture is stirred 30 minutes.Added in room temperature 3- (pyridine -2- bases) -2H- pyridos [1,2-a] [1,3,5] triazine -2,4 (3H)-diketone (0.406g, 1.690mmol), then Stirred 24 hours at 65 DEG C.Reactant mixture is cooled to room temperature, then distributed in icy water (30mL) and EtOAc Between (100mL).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain thick material, it is palm fibre Color solid (TLC eluant, eluents：70%EtOAc/ hexanes:R_f-0.3；UV activation).Crude residue is purified through column chromatography, is used Neutral alumina simultaneously uses 20%EtOAc/ Hex, obtains pure (4S) -9- methyl -7- (6- picoline -3- bases)-N- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (75.7mg, 0.193mmol, 17.15% yield), it is pale solid, LCMS (m/z):387.3[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.59-14.03 (m, 1H), 9.07 (d, J=2.19Hz, 1H), 8.59 (dd, J=8.22,2.52Hz, 1H), 8.30-8.51 (m, 1H), 8.18 (d, J=8.55Hz, 1H), 7.58-7.83 (m, 1H), 7.28-7.40 (m, 2H), 6.91-7.05 (m, 1H), 5.69 (dd, J=5.92,3.07Hz, 1H), 3.06-3.23 (m, 3H), 2.94-3.05(m,1H),2.63(s,3H),2.49(s,3H),2.24-2.38(m,1H),1.99-2.12(m,1H)

Embodiment 10

Synthesize (4S) -8- cyano group -7- (6- picoline -3- bases)-N- (pyridin-3-yl) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaIn the solution of the stirring of -8- formonitrile HCNs (200mg, 0.721mmol) in THF (15ml, in seal pipe) Triphosgene (128mg, 0.433mmol) is added, is then stirred to room temperature, continues 1h.Be subsequently added pyridine -3- amine (102mg, 1.082mmol) with DIPEA (280mg, 2.164mmol), 16h are heated at 75 DEG C.Reactant mixture is poured into saturation NaHCO₃It is molten In liquid (50mL) and it is extracted with ethyl acetate (3x50mL).The organic layer of merging is through anhydrous Na₂SO₄Dry and be concentrated under reduced pressure, obtain Crude product.Crude mixture purifies (silica gel through column chromatography：100-200 mesh, uses the gradient mixture of 1% ethanol/methylene It is used as eluant, eluent), obtain (4S) -8- cyano group -7- (6- picoline -3- bases)-N- (pyridin-3-yl) -3,4- dihydros-Isosorbide-5-Nitrae-bridge Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (140mg, 0.350mmol, 48.5% yield), its For pale solid (TLC systems:R_f:0.3, eluant, eluent：5% ethanol/methylene), LCMS (m/z):398.29[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 12.58 (s, 1H), 8.98 (d, J=2.19Hz, 1H), 8.44 (d, J= 2.41Hz, 1H), 8.31 (dd, J=4.71,1.43Hz, 1H), 8.12-8.02 (m, 2H), 7.84 (s, 1H), 7.38 (d, J= 8.11Hz, 1H), 7.29-7.24 (m, 1H), 5.71 (dd, J=5.92,2.85Hz, 1H), 3.33-3.21 (m, 2H), 3.02- 3.17(m,2H),2.69(s,3H),2.45-2.32(m,1H),2.22-2.08(m,1H)。

Embodiment 11

At 0 DEG C to the bromo- 7- of (4S) -8- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diazaNaH is added in the solution of the stirring of (300mg, 0.906mmol) in THF (20mL) (65.2mg,2.72mmol).Then by reactant mixture 30 DEG C stir 1h, add 3- (pyridine -2- bases) -2H- pyridos [1, 2-a] [1,3,5] triazine -2,4 (3H)-diketone (435mg, 1.812mmol), then stirs 15h by reactant mixture at 70 DEG C. Reactant mixture is poured into cold water (10mL) and is extracted with ethyl acetate (2x20mL).The organic layer of merging is through anhydrous sodium sulfate Dry and be concentrated under reduced pressure, obtain crude product.Crude mixture is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent： 3%MeOH/EtOAc), the bromo- 7- of (4S) -8- (2- picoline -4- bases)-N- (pyridine -2- bases) -3,4- dihydro-Isosorbide-5-Nitraes-are obtained Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (290mg, 0.639mmol, 70.5% yield), It is pale solid (TLC systems:R_f- 0.3, eluant, eluent：5% ethanol/methylene), LCMS (m/z):451.24[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 12.85 (s, 1H), 8.63 (d, J=5.26Hz, 1H), 8.30-8.25 (m,1H),8.15-8.09(m,1H),7.82(s,1H),7.76(s,1H),7.70-7.63(m,2H),7.02–6.93(m,1H), 5.66 (dd, J=5.92,3.07Hz, 1H), 3.38-3.09 (m, 3H), 3.01 (dd, J=12.06,3.29Hz, 1H), 2.72 (s, 3H), 2.34 (dddd, J=14.20,10.08,6.08,3.84Hz, 1H), 2.16-2.02 (m, 1H).

Embodiment 12

Synthesize (4S) -8- bromo- 7- (2- picoline -4- bases)-N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the bromo- 7- of (4S) -8- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diazaTriphosgene is added in the solution of the stirring of (250mg, 0.755mmol) in THF (10mL) (134mg,0.453mmol).Then by reactant mixture 30 DEG C stir 30min and add pyridine -3- amine (142mg, 1.510mmol).Reactant mixture is stirred into 15h at 70 DEG C.Allow to cool to RT and reactant mixture is poured into cold water (20mL) In and be extracted with ethyl acetate (2X20mL).The organic layer of merging is through anhydrous Na₂SO₄Dry and be concentrated under reduced pressure, obtain rough chemical combination Thing.Crude mixture is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：3%MeOH/EtOAc), 500mg is obtained, It is 90% to determine purity by LCMS.Half pure compound purifies (post through preparation HPLC：XS PHENYL HEXYL (250X4.6mm, 5 μ), mobile phase:A:0.01M ammonium hydrogen carbonate B:ACN, gradient：Time/%B:0/10,1/10,10/50,15/ 50,18/98,20/98,20.1/10,25/10, column temperature：Environment temperature, flow velocity:1.0ml/min, diluent：ACN), obtain The bromo- 7- of (4S) -8- (2- picoline -4- bases)-N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza- 5 (2H)-formamides (175mg, 0.387mmol, 51.3% yield), it is pale solid (TLC: 10%MeOH/ ethyl acetate, R_f:0.3), LCMS (m/z):451.21[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 12.56 (s, 1H), 8.69 (d, J=5.26Hz, 1H), 8.43 (d, J= 2.63Hz, 1H), 8.28 (dd, J=4.71,1.42Hz, 1H), 8.02-7.94 (m, 1H), 7.83 (s, 1H), 7.49-7.41 (m, 2H), 7.21 (dd, J=8.33,4.82Hz, 1H), 5.67 (dd, J=5.81,3.18Hz, 1H), 3.38-3.11 (m, 3H), 3.05-2.98 (m, 1H), 2.69 (s, 3H), 2.35 (dddd, J=14.20,10.03,6.03,3.95Hz, 1H), 2.15-2.01 (m,1H)。

Embodiment 13

Synthesize (4S) -9- methyl -7- (6- picoline -3- bases)-N- (pyridin-3-yl) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

In a nitrogen atmosphere in room temperature by N, N- dimethyl aminopyridines (0.482g, 3.94mmol) add to (4S) -9- first Base -7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza (0.350g, 1.314mmol) and pyridin-3-yl phenyl carbamate (0.845g, 3.94mmol) are in tetrahydrofuran (20mL) Stirring solution in.Reactant mixture is stirred 48 hours at 65 DEG C.Reactant mixture is cooled to room temperature and is removed under reduced pressure Solvent, is then distributed between water (20mL) and EtOAc (70mL).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filtering And evaporate filtrate, thick material is obtained, it is brown solid (TLC eluant, eluents：100%EtOAc:R_f-0.3；UV activation).Add Plus ethyl acetate is into reactive material and stirs 10 minutes.Filtering reactive material is simultaneously washed with EtOAc.Filtrate is concentrated, obtained Brown solid (TLC eluant, eluents：100%EtOAc:R_f-0.2；UV activation).Thick material is purified through column chromatography, uses neutral oxygen Change aluminium and use 60%EtOAc/ Hex, obtain pure (4S) -9- methyl -7- (6- picoline -3- bases)-N- (pyridine -3- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (0.115g, 0.296mmol, 22.56% yield), it is white solid, LCMS (m/z) 387.3 [M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.29 (s, 1H), 8.92 (d, J=1.97Hz, 1H), 8.53 (d, J= 2.19Hz, 1H), 8.28 (br d, J=3.51Hz, 1H), 8.19 (br d, J=8.33Hz, 1H), 7.99 (dd, J=8.00, 2.30Hz, 1H), 7.32 (d, J=7.89Hz, 1H), 7.22-7.29 (m, 1H), 7.19 (s, 1H), 5.68 (dd, J=5.92, 3.07Hz, 1H), 3.17 (t, J=7.34Hz, 2H), 3.07-3.13 (m, 1H), 2.97-3.05 (m, 1H), 2.64 (s, 3H), 2.50 (s, 3H), 2.32 (td, J=13.76,6.47Hz, 1H), 1.99-2.14 (m, 1H)

Embodiment 14

Synthesize (4S) -8- fluoro- 7- (2- picoline -4- bases)-N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

By K₃PO₄(515mg, 2.427mmol) adds to the chloro- 8- of (4S) -7- fluoro- N- (pyridin-3-yl) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (405mg, 1.213mmol), (2- picolines- 4- yls) in solution of the boric acid (216mg, 1.578mmol) in 1,4- dioxanes (20mL) and water (3mL).By reactant mixture Deaerate 15min, then adds Pd in room temperature₂(dba)₃(55.6mg, 0.061mmol) and X-phos (57.8mg, 0.121mmol), And reactant mixture is heated into 3h at 80 DEG C.Reactant mixture is cooled to room temperature and distributed in water (25mL) and EtOAc Between (60mL).The organic layer of separation is through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into crude product.Crude compound Purified (silica gel：100-200 mesh, eluant, eluent：90% ethyl acetate/hexane), obtain the fluoro- 7- of (4S) -8- (2- picolines - 4- yls)-N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first Acid amides (195mg, 0.487mmol, 40.1% yield), it is pale solid (TLC eluant, eluents：Net ethyl acetate R_f:0.2), LCMS(m/z):391.30[M+H]⁺。

¹H NMR(400MHz,CDCl₃-d):δ ppm 12.7 (s, 1H), 8.68 (d, J=4.8Hz, 1H), 8.58 (d, J= 2.4Hz, 1H), 8.31 (dd, J=1.6Hz, 4.8Hz, 1H), 8.11-8.07 (m, 1H), 7.60 (s, 1H), 7.55-7.53 (m, 1H), 7.45 (d, J=10.4Hz, 1H), 7.27-7.24 (m, 1H), 5.70-5.68 (m, 1H), 3.33-3.14 (m, 3H), 3.05-3.01(m,1H),2.68(s,3H),2.38-2.30(m,1H),2.12-2.06(m,1H)。

Embodiment 15

Synthesize (4S) -8- fluoro- 7- (2- picoline -4- bases)-N- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

By K₃PO₄(445mg, 2.097mmol) adds to the fluoro- N- of the chloro- 8- of (4S) -7- (pyridine -2- bases) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (350mg, 1.049mmol), (2- picolines- 4- yls) stirring of the boric acid (187mg, 1.363mmol) in 1,4- dioxanes (10mL) and water (2mL) solution in.Will reaction Mixture degassing 15min, adds Pd₂(dba)₃(48.0mg, 0.052mmol) and x-phos (50.0mg, 0.105mmol).Will be anti- Mixture is answered to stir 16h at 80 DEG C.By reactant mixture be cooled to room temperature and distribute water (25mL) and EtOAc (60mL) it Between.The organic layer of separation is through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain crude compound.Crude compound is through fast Fast column chromatography purifies (silica gel：100-200 mesh, eluant, eluent：90% ethyl acetate/hexane), obtain (4S) -8- fluoro- 7- (2- methyl Pyridin-4-yl)-N- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (120mg, 0.306mmol, 29.2% yield), it is pale solid (TLC eluant, eluents：Net ethyl acetate R_f:.0.4), LCMS (m/z):391.27[M+H]⁺

¹H NMR(400MHz,CDCl₃-d):δ ppm 13.05 (s, 1H), 8.63 (d, J=5.6Hz, 1H), 8.35-8.33 (m, 1H), 8.19 (d, J=8.4Hz, 1H), 8.09 (s, 1H), 7.81-7.80 (m, 1H), 7.71-7.67 (m, 1H), 7.43 (d, J=10.4Hz, 1H), 7.25-6.99 (m, 1H), 5.71-5.68 (m, 1H), 3.32-3.14 (m, 3H), 3.03-2.99 (m, 1H),2.74(s,3H),2.36-2.32(m,1H),2.09-2.05(m,1H).

Embodiment 16

Synthesize (4S) -8- chloro- 7- (6- picoline -3- bases)-N- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

(4S) -8- chloro- 7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- bridges to be stirred at 0 DEG C sub- under a nitrogen Picoline simultaneously [2,3-b] [1,4] diaza(350mg, 1.221mmol), NaH (43.9mg, 1.831mmol) are in tetrahydrochysene furan The solution muttered in (THF) (20mL).Then reactant mixture is stirred into 30min at 30 DEG C and adds 3- (pyridine -2- bases) -2H- Pyrido [1,2-a] [1,3,5] triazine -2,4 (3H)-diketone (352mg, 1.465mmol).By reactant mixture in 70 DEG C of stirrings 16h.Reactant mixture is poured into cold water (50mL) and is extracted with ethyl acetate (2x75mL).Organic layer is dry through anhydrous magnesium sulfate It is dry, filter and be concentrated in vacuo, obtain crude compound.Crude mixture is through flash column chromatography (silica gel：100-200 mesh, is washed De- agent：3%MeOH/EtOAc), 500mg is obtained, it is 90% to determine purity by LCMS.Half pure compound is through preparation HPLC Purify (post：XS PHENYL HEXYL (250X4.6mm, 5 μ), mobile phase:A:0.01M ammonium hydrogen carbonate B:ACN, gradient：When Between/%B:0/10,1/10,10/50,15/50,18/98,20/98,20.1/10,25/10, column temperature：Environment temperature, flow velocity: 1.0ml/min, diluent：ACN), the chloro- 7- of (4S) -8- (6- picoline -3- bases)-N- (pyridine -2- bases) -3,4- bis- is obtained Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamide (240mg, 0.589mmol, 48.3% Yield), it is pale solid (TLC systems；R_f- 0.3,5% ethanol/methylene), LCMS (m/z):407.29[M+H]⁺。

¹H NMR(400MHz,CDCl₃) δ ppm 12.93 (s, 1H), 9.07 (d, J=2.19Hz, 1H), 8.28 (ddd, J= 4.82,1.97,0.88Hz, 1H), 8.21 (dd, J=8.11,2.41Hz, 1H), 8.11-8.06 (m, 1H), 7.68-7.63 (m, 2H), 7.33 (d, J=8.11Hz, 1H), 6.96 (ddd, J=7.29,4.88,0.99Hz, 1H), 5.66 (dd, J=6.03, 3.18Hz, 1H), 3.35-3.12 (m, 3H), 3.01 (dd, J=12.28,3.29Hz, 1H), 2.63 (s, 3H), 2.33 (dddd, J =14.14,10.03,5.86,3.73Hz, 1H), 2.09 (dt, J=14.20,7.04Hz, 1H).

Embodiment 17

Synthesize (4S) -8- methyl -7- (2- picoline -4- bases)-N- (pyridin-3-yl) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4S) -8- methyl -7- (2- picoline -4- the bases) -2,3,4,5- four for stirring 30min at 20 DEG C under a nitrogen Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(300mg, 1.126mmol), triphosgene (201mg, 0.676mmol), added in solution of the DIPEA (728mg, 5.63mmol) in tetrahydrofuran (THF) (10mL) solid pyridine- 3- amine (159mg, 1.690mmol).Reactant mixture is stirred into 16h at 65 DEG C.By reactant mixture saturation NaHCO₃Solution Dilute and be extracted with ethyl acetate (3 × 150mL).Organic layer is through anhydrous Mg₂SO₄Dry, filter, be concentrated in vacuo, obtain roughization Compound.Crude product is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：3%MeOH/EtOAc), (4S) -8- is obtained Methyl -7- (2- picoline -4- bases)-N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (200mg, 0.499mmol, 44.3% yield), it is pale solid.(TLC systems:- R_f:- 0.5,10% methanol/ethyl acetate), LCMS (m/z):387.31[M+H]⁺。

¹H NMR(400MHz,CDCl₃) δ ppm 13.01 (s, 1H), 8.67 (d, J=5.26Hz, 1H), 8.45 (d, J= 2.41Hz, 1H), 8.26 (dd, J=4.82,1.32Hz, 1H), 8.09-7.94 (m, 1H), 7.47 (s, 1H), 7.34 (s, 1H), 7.20 (dd, J=8.33,4.60Hz, 1H), 5.66 (dd, J=5.92,3.07Hz, 1H), 3.35-3.11 (m, 3H), 3.00 (dd, J=12.28,3.29Hz, 1H), 2.68 (s, 3H), 2.38-2.28 (m, 4H), 2.11-1.98 (m, 1H), 2.11-1.98 (m,1H)。

Embodiment 18

Synthesize (4S) -8- chloro- N- (2- (((R)-tetrahydrofuran -3- bases) epoxide) pyrimidine-4-yl) -7- (3- (fluoroforms Base) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diazaIn the solution of the stirring of (300mg, 0.883mmol) in THF (15mL, in seal pipe) Triphosgene (157mg, 0.530mmol), and stirring 30min are added, TEA (0.738mL, 5.30mmol) and (R) -2- is then added ((tetrahydrofuran -3- bases) epoxide) pyrimidine -4- amine (192mg, 1.060mmol), then stirs 16h at 80 DEG C.(TLC eluant, eluents： 10%MeOH/EtOAc, Rf:0.6).Reactant mixture is cooled to room temperature；THF is distilled out, then distributed at water (25mL) Between EtOAc (40mL).Separate organic layer and through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain rough chemical combination Thing.Crude compound is through flash column chromatography (silica gel:100-200 mesh, eluant, eluent：1% methanol/ethyl acetate), it must expire The chloro- N- of product (4S) -8- (2- (((R)-tetrahydrofuran -3- bases) epoxide) pyrimidine-4-yl) -7- (3- (trifluoromethyl) benzene of prestige Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (110mg, 0.195mmol, 22.12% yield), it is pale solid.LCMS(m/z):547.10[M+H]⁺, Rt=2.65min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.02 (s, 1H), 8.41 (d, J=5.48Hz, 1H), 8.08-7.97 (m, 2H), 7.95-7.86 (m, 1H), 7.86-7.73 (m, 2H), 7.73-7.57 (m, 1H), 5.46 (dd, J=5.92, 3.07Hz, 1H), 4.92 (dd, J=4.38,1.53Hz, 1H), 3.76 (q, J=7.60Hz, 1H), 3.69-3.59 (m, 1H), 3.54 (dd, J=10.52,1.75Hz, 1H), 3.39 (dd, J=10.41,4.71Hz, 1H), 3.29 (s, 1H), 3.25-3.07 (m, 2H), 3.07-2.79 (m, 1H), 2.41-2.13 (m, 1H), 1.99 (t, J=6.69Hz, 1H), 1.91-1.63 (m, 2H).

Embodiment 19

Synthesize (4S) -8- chloro- N- (2- (((S)-tetrahydrofuran -3- bases) epoxide) pyrimidine-4-yl) -7- (3- (fluoroforms Base) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaThree are added in the solution of the stirring of (400mg, 1.177mmol) in THF (10mL, in seal pipe) Phosgene (349mg, 1.177mmol) and TEA (0.985mL, 7.06mmol) stir 30min in room temperature, then add (S) -2- ((tetrahydrofuran -3- bases) epoxide) pyrimidine -4- amine (427mg, 2.355mmol), then heats 16h at 80 DEG C.(TLC eluant, eluents： Net EtOAc, Rf:0.2).Reactant mixture is cooled to room temperature；THF is distilled out, then distributed in water (25mL) and EtOAc Between (2x30mL).Separate organic layer and through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain crude compound.It is rough Compound is through flash column chromatography (silica gel:100-200 mesh, eluant, eluent：90%EtOAc/ hexanes), obtain desired product The chloro- N- of (4S) -8- (2- (((S)-tetrahydrofuran -3- bases) epoxide) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (150mg, 0.272mmol, 23.10% yield), it is pale solid.LCMS(m/z):547.07[M+H]⁺, Rt=2.62min.

1H NMR(400MHz,CDCl₃):δ ppm 12.97 (s, 1H), 8.34 (d, J=5.70Hz, 1H), 8.12-7.92 (m, 2H), 7.87-7.46 (m, 4H), 5.65 (dd, J=5.92,3.07Hz, 1H), 5.07 (ddt, J=6.77,4.63,2.19, 2.19Hz, 1H), 4.07-3.46 (m, 4H), 3.38-3.09 (m, 3H), 3.03 (dd, J=12.28,3.29Hz, 1H), 2.36 (dddd, J=14.22,10.06,6.08,4.06Hz, 1H), 2.16-1.97 (m, 2H), 1.94-1.81 (m, 1H).

Embodiment 20

Synthesis (4S) -8- chloro- 7- (2- picoline -4- bases)-N- (2- (((S)-tetrahydrofuran -3- bases) epoxide) pyrimidine - 4- yls) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to the chloro- 7- of (4S) -8- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diazaAdd in the solution of the stirring of (300mg, 1.046mmol) in THF (15mL, in seal pipe) Enter triphosgene (310mg, 1.046mmol) and TEA (0.875mL, 6.28mmol), and stirring 30min, then add (S) -2- ((tetrahydrofuran -3- bases) epoxide) pyrimidine -4- amine (379mg, 2.092mmol), then heats 16h at 80 DEG C.(TLC eluant, eluents： Net EtOAc, Rf:0.3).Reactant mixture is cooled to room temperature；THF is distilled out, then distributed in water (25mL) and EtOAc Between (2x30mL).Separate organic layer and through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain crude compound.It is rough Compound is through flash column chromatography (silica gel:100-200 mesh, eluant, eluent：90%EtOAc/ hexanes), obtain half pure compound And (condition is further purified by preparation HPLC：Post：XBridge C 18 (75X4.6mm, 3.5 μ) mobile phase:A:0.01M Ammonium hydrogen carbonate B:ACN:Gradient：Time/%B:0/5,0.8/5,5/50,8/95,12/95,12.1/5,15/5:Column temperature：Environment Temperature, flow velocity:1.0ml/min:Diluent：ACN), obtain the chloro- 7- of desired product (4S) -8- (2- picoline -4- bases) - N- (2- (((S)-tetrahydrofuran -3- bases) epoxide) pyrimidine-4-yl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (110mg, 0.221mmol, 21.13% yield), it is pale solid.LCMS (m/z):494.04[M+H]⁺, Rt=1.67min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.93 (s, 1H), 8.68 (d, J=5.26Hz, 1H), 8.33 (d, J= 5.48Hz, 1H), 7.73 (d, J=5.70Hz, 1H), 7.68 (s, 1H), 7.63 (s, 1H), 7.59 (s, 1H), 5.63 (dd, J= 5.92,3.07Hz, 1H), 5.19 (dq, J=5.97,3.05Hz, 1H), 4.01-3.88 (m, 2H), 3.85-3.72 (m, 2H), 3.37-3.21 (m, 2H), 3.18-3.11 (m, 1H), 3.01 (dd, J=12.28,3.07Hz, 1H), 2.73 (s, 3H), 2.42- 2.29(m,1H),2.11-2.05(m,3H)。

Embodiment 21

Synthesis (4S) -8- chloro- 7- (2- picoline -4- bases)-N- (2- ((tetrahydrochysene -2H- pyrans -4- bases) epoxide) pyrimidine - 4- yls) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to the chloro- 7- of (4S) -8- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diazaAdd in the solution of the stirring of (300mg, 1.046mmol) in THF (10mL, in seal pipe) Enter triphosgene (155mg, 0.523mmol), and stirring 30min, then add DIPEA (0.914mL, 5.23mmol) and 2- ((four Hydrogen -2H- pyrans -4- bases) epoxide) pyrimidine -4- amine (408mg, 2.092mmol), then stirs 16h at 80 DEG C.(TLC systems: Rf-0.2,EtOAc).Reactant mixture is cooled to room temperature and THF is distilled out, by crude product distribution in water (25mL) and EtOAc Between (2x40mL).The organic layer of separation is through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain crude compound.It is rough Compound is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：80% ethyl acetate/hexane), obtain 57% pure thing Matter, therefore it is purified into (condition again by preparation HPLC：Post：XS PHENYL HEXYL (250X4.6mm, 5 μ), flowing Phase:A:0.01M ammonium hydrogen carbonate B:ACN, gradient：Time/%B:0/10,1/10,10/50,15/50,18/98,20/98,20.1/ 10,25/10 column temperature：Environment temperature, flow velocity:0.8ml/min, diluent：ACN), the chloro- 7- of desired product (4S) -8- are obtained (2- picoline -4- bases)-N- (2- ((tetrahydrochysene -2H- pyrans -4- bases) epoxide) pyrimidine-4-yl) -3,4- dihydro -1,4- bridges are sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (96.6mg, 0.183mmol, 17.45% yield), its For faint yellow solid.LCMS(m/z):508.1[M+H]⁺, Rt=1.75min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.92 (s, 1H), 8.68 (d, J=5.26Hz, 1H), 8.35 (d, J= 5.70Hz, 1H), 7.73 (d, J=5.48Hz, 1H), 7.68 (s, 1H), 7.63 (s, 1H), 7.59 (d, J=4.82Hz, 1H), 5.67-5.60 (m, 1H), 4.92 (dt, J=8.22,4.22Hz, 1H), 4.03-3.86 (m, 2H), 3.56-3.45 (m, 2H), 3.37-3.22 (m, 2H), 3.19-3.11 (m, 1H), 3.02 (dd, J=12.17,3.18Hz, 1H), 2.72 (s, 3H), 2.40- 2.29 (m, 1H), 2.07 (dt, J=14.52,7.54Hz, 1H), 1.94 (br s, 2H), 1.82-1.70 (m, 2H).

Embodiment 22

Synthesis (4S) -8- chloro- 7- (2- picoline -4- bases)-N- (2- (((R)-tetrahydrofuran -3- bases) epoxide) pyrimidine - 4- yls) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C under a nitrogen to the chloro- 7- of (4S) -8- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diazaAdded in the solution of the stirring of (200mg, 0.697mmol) in THF (20mL) Triethylamine (0.486mL, 3.49mmol) and triphosgene (103mg, 0.349mmol), are then stirred at room temperature 30min.It is anti-to this Addition (R) -2- ((tetrahydrofuran -3- bases) epoxide) pyrimidine -4- amine (164mg, 0.907mmol) in mixture is answered, then 80 DEG C stirring 15h.(TLC systems:5% methanol/ethyl acetate .Rf values：0.5.).(10mL) and use is diluted with water in reactant mixture Ethyl acetate extracts (2x15mL).(10mL) is washed with water in the organic layer of merging, through anhydrous sodium sulfate drying and is concentrated under reduced pressure, and obtains Obtain crude compound.Crude product is obtained through flash column chromatography (100-200 silica gel is eluted with 1%MeOH/ ethyl acetate) The chloro- 7- of desired product (4S) -8- (2- picoline -4- bases)-N- (2- (((R)-tetrahydrofuran -3- bases) epoxide) pyrimidine -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (120mg, 0.237mmol, 33.9% yield), it is pale pink solid.LCMS(m/z):494.07[M+H]⁺, Rt=1.69min.

¹H NMR(400MHz,CDCl3):δ ppm 12.98 (s, 1H), 8.66 (d, J=5.04Hz, 1H), 8.34 (d, J= 5.70Hz, 1H), 7.74 (d, J=5.48Hz, 1H), 7.68 (s, 1H), 7.58 (s, 1H), 7.55-7.52 (m, 1H), 5.64 (dd, J=5.81,3.18Hz, 1H), 5.17 (tt, J=5.26,2.52Hz, 1H), 3.97-3.85 (m, 2H), 3.77-3.65 (m, 2H), 3.35-3.11 (m, 3H), 3.03 (dd, J=12.28,3.07Hz, 1H), 2.71 (s, 3H), 2.42-2.28 (m, 1H),2.15-1.96(m,3H)。

Embodiment 23

Synthesize (4S) -8- chloro- N- (5- ((R) -2,3- dihydroxy propoxyl group) pyridin-3-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (5- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridin-3-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaHCl/water solution is added in the solution of stirring of -5 (the 2H)-formamides (200mg, 0.339mmol) in methanol (10mL) (1.0ml, 32.9mmol), lasts 10min, then stirs 1h at 0 DEG C.(TLC eluant, eluents：10% methanol/DCM, Rf:0.1；UV Activation).Reactive material is concentrated under reduced pressure and gained viscous brown oily thing is dissolved in water, saturated sodium bicarbonate aqueous solution is used (10mL) alkalizes, and is then extracted with 10% methanol/DCM (40mL).Organic layer is dried over sodium sulfate with salt water washing (20mL), Filter and be concentrated under reduced pressure, obtain the chloro- N- of (4S) -8- (5- ((R) -2,3- dihydroxy propoxyl group) pyridin-3-yl) -7- (3- (trifluoros Methyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (120mg, 0.217mmol, 64.1% yield), it is pale solid.LCMS(m/z):550.09[M+H]⁺, Rt= 2.03min。

¹H NMR(400MHz,DMSO-d₆):δppm 12.55(s,1H),8.02-8.18(m,2H),7.75-8.01(m, 5H), 7.62 (s, 1H), 5.47 (br s, 1H), 5.00 (br s, 2H), 4.00 (br dd, J=9.21,3.73Hz, 1H), 3.73-3.92(m,2H),3.34-3.60(m,2H),2.83-3.25(m,4H),2.12-2.30(m,1H),1.83-2.09(m, 1H)。

Embodiment 24

Synthesize (4S) -8- chloro- N- (6- ((R) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (6- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3b] [1,4] diaza4M is added in the solution of stirring of -5 (the 2H)-formamides (0.25g, 0.423mmol) in DCM (5mL) and methanol (10mL) HCl dioxane solutions (1.586mL, 6.35mmol), then stir 4h at 0 DEG C.(TLC eluant, eluents：10%MeOH/EtOAc: R_f-0.3；UV activation).By reactant mixture by adding saturated sodium bicarbonate solution alkalization (until pH-8-9), Ran Hounong Contracting volatile matter.By residue diluted with water (10mL) and it is extracted into ethyl acetate (2x25mL).The organic extract warp of merging Anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain crude product.Thick material purifies (neutral alumina, elution through column chromatography Agent：50% ethyl acetate/hexane), obtaining the chloro- N- of desired product (4S) -8-, (6- ((R) -2,3- dihydroxy propoxyl group) is phonetic Pyridine -4- bases) -7 (3 (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas-5 (2H)-formamide, it is white solid.LCMS(m/z):551.03[M+H]⁺,R_t=2.35min.

¹H NMR(400MHz,CDCl₃):δppm 13.14(s,1H),8.34(s,1H),8.16(s,1H),8.08(br d, J=7.67Hz, 1H), 7.77-7.58 (m, 3H), 7.53 (s, 1H), 5.63 (br s, 1H), 4.57-4.36 (m, 2H), 4.03 (br s, 1H), 3.69 (br d, J=12.93Hz, 2H), 3.44 (br s, 1H), 3.30-3.03 (m, 3H), 2.97-2.63 (m, 1H), 2.49 (br s, 1H), 2.35 (br dd, J=9.76,4.93Hz, 1H), 2.11-2.00 (m, 1H).

Embodiment 25

Synthesize (4S) -8,9- dimethyl -7- (2- picoline -4- bases)-N- (pyridine -2- bases) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Under a nitrogen at 0 DEG C to (4S) -8,9- dimethyl -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysenes -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diazaAdded in the solution of (250mg, 0.892mmol) in THF (25mL) NaH (214mg, 5.35mmol), is then stirred at room temperature 30min.Then 3- (pyridine -2- bases) -2H- pyridos [1,2- is added A] [1,3,5] triazine -2,4 (3H)-diketone (321mg, 1.338mmol) and by reactant mixture 65 DEG C heat 16h.(TLC is washed De- agent：70%EtOAc/ hexanes:R_f-0.4；UV activation).By reactant mixture be cooled to room temperature and distribute in water (30mL) and Between EtOAc (100mL).Organic layer is separated, through anhydrous Na₂SO₄Dry, filter and concentrate filtrate, obtain crude product.Thick thing Matter purifies (neutral alumina, eluant, eluent through column chromatography：50% ethyl acetate/hexane), obtain desired product (4S) -8,9- bis- Methyl -7- (2- picoline -4- bases)-N- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (0.164g, 0.403mmol, 45.1% yield), it is white solid LCMS (m/z): 401.15[M+H]⁺,R_t=1.55min.

¹H-NMR(400MHz,CDCl₃):δ ppm 13.43 (s, 1H), 8.59 (d, J=5.26Hz, 1H), 8.26 (dt, J= 4.82,0.99Hz, 1H), 8.12 (d, J=8.55Hz, 1H), 7.67-7.53 (m, 2H), 7.32 (dd, J=5.26,1.10Hz, 1H), 6.93 (ddd, J=7.29,4.88,0.99Hz, 1H), 5.63 (dd, J=5.92,3.07Hz, 1H), 3.22-3.05 (m, 3H),3.05-2.96(m,1H),2.70(s,3H),2.46(s,3H),2.36-2.24(m,4H),2.12-1.97(m,1H)

Embodiment 26

Synthesize (4S) -8- chloro- N- (4- phenyl pyrimidine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

It is sub- to the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- bridges in room temperature under a nitrogen Picoline simultaneously [2,3-b] [1,4] diazaAdd in the solution of the stirring of (350mg, 1.030mmol) in THF (35mL) Enter triethylamine (0.862mL, 6.18mmol), triphosgene (306mg, 1.030mmol) and stir 30min.Then 4- phenyl is added Reactant mixture is simultaneously heated 16h by pyrimidine -2- amine (176mg, 1.030mmol) at 65 DEG C.(TLC eluant, eluents：100%EtOAc: R_f-0.2；UV activation).Reactant mixture is cooled to room temperature, concentrated, residue is distributed in water (30mL) and DCM (100mL) Between.Organic layer is separated, through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain crude compound.Thick material is through post color Spectrum purifying (neutral alumina, eluant, eluent：70% ethyl acetate/hexane), obtain the chloro- N- of (4S) -8- (4- phenyl pyrimidines -2- Base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (262mg, 0.488mmol, 47.4% yield), it is pale solid.LCMS(m/z):537.13[M+H ]⁺,R_t=2.84min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.27 (s, 1H), 8.65 (d, J=5.26Hz, 1H), 8.13 (s, 1H), 8.05 (d, J=7.89Hz, 1H), 7.89-7.81 (m, 2H), 7.72 (d, J=7.89Hz, 1H), 7.68 (s, 1H), 7.58- 7.46 (m, 2H), 7.44-7.35 (m, 3H), 5.78 (dd, J=5.81,3.18Hz, 1H), 3.38-3.13 (m, 3H), 3.03 (dd, J=12.17,3.18Hz, 1H), 2.43-2.31 (m, 1H), 2.19-2.07 (m, 1H).

Embodiment 27

Synthesize (4S) -8- chloro- N- (4- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (4- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaHCl/water solution is added in the solution of stirring of -5 (the 2H)-formamides (450mg, 0.838mmol) in methanol (10mL) (0.509mL, 16.76mmol, 36%), then stirs 2h at 0 DEG C.(TLC eluant, eluents：100%EtOAc:R_f-0.2；UV is activated ).Reactant mixture is alkalized (until pH-8-9) with saturated sodium bicarbonate solution and solvent is evaporated under reduced pressure.Residue is distributed Between water (10mL) and dichloromethane (2x20mL).Organic layer is separated through anhydrous sodium sulfate drying, is filtered and by filter vacuum Evaporation, obtains crude compound.Thick material purifies (neutral alumina, eluant, eluent through column chromatography：5%MeOH/DCM), institute is obtained The chloro- N- of product (4S) -8- (4- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (2- picoline -4- bases) -3 is needed, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (195mg, 0.389mmol, 46.4% yield), it is faint yellow solid.LCMS(m/z):497.10[M+H]⁺,R_t=1.24min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.91 (s, 1H), 8.63 (d, J=5.48Hz, 1H), 8.08 (d, J= 5.70Hz, 1H), 7.80 (s, 1H), 7.76 (d, J=2.19Hz, 1H), 7.68 (dd, J=5.15,1.21Hz, 1H), 7.65 (s, 1H), 6.69-6.46 (m, 1H), 5.63 (dd, J=5.92,3.07Hz, 1H), 4.13 (s, 3H), 3.87-3.81 (m, 1H), 3.79-3.72 (m, 1H), 3.36-3.13 (m, 3H), 3.01 (dd, J=12.28,3.29Hz, 1H), 2.71 (s, 4H), 2.41- 2.28(m,1H),2.20-2.03(m,2H)。

Embodiment 28

Synthesize (4S) -8- chloro- N- (2- ((S) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-carboxamide hydrochlorides

At 0 DEG C to the chloro- N- of (4S) -8- (2- ((S) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (3- (fluoroforms Base) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (200mg, 2M HCl diethyl ether solution (5mL, 165mmol) 0.363mmol) is added in the solution in methanol (5mL), is then stirred in room temperature Mix 2h.(TLC eluant, eluents：15%MeOH/DCM:R_f=0.2.；UV activation).Reactant mixture is concentrated in vacuo and by remnants Thing grinds (2x10mL) with pentane, obtains the chloro- N- of (4S) -8- (2- ((S) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine hydrochlorate (160mg, 0.270mmol, 74.4% yield), it is pale solid.LCMS(m/z):551.10[M+H]⁺,R_t =2.17min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 12.95 (s, 1H), 8.44 (d, J=5.70Hz, 1H), 8.13-8.08 (m, 1H), 8.06 (s, 1H), 7.98 (s, 1H), 7.90-7.83 (m, 2H), 7.68 (d, J=5.70Hz, 1H), 5.49 (dd, J= 6.03,2.96Hz,1H),3.99-3.85(m,2H),3.75-3.68(m,1H),3.43-3.32(m,3H),3.28-3.17(m, 2H), 3.11 (dd, J=11.73,3.18Hz, 1H), 2.35-2.23 (m, 1H), 2.04 (dt, J=13.98,6.93Hz, 1H).

Embodiment 29

Synthesize (4S) -8- chloro- N- (5- ((R) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to the chloro- N- of (4S) -8- (5- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaAdded in solution of -5 (the 2H)-formamides (220mg, 0.409mmol) in methanol (5mL) HCl/water solution (5mL, 10.00mmol) and stir 3h.(5% methanol of TLC systems/DCM.Rf values：0.1).Methanol in vacuo is removed and at 0 DEG C by remnants Thing saturation NaHCO₃It is basified, and water layer is extracted (2x20mL) with DCM.Organic layer is separated through anhydrous Na₂SO₄Dry, mistake Filter and concentrate, obtain crude product.Crude compound obtains the chloro- N- of (4S) -8- (5- ((R) -2,3- bis- with triturated under ether (5mL) Hydroxy propyloxy group) pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (145mg, 0.282mmol, 69.0% yield), it is faint yellow solid.LCMS(m/ z):498.14[M+H]⁺, Rt=1.44min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.72-13.12 (m, 1H), 8.92 (d, J=1.32Hz, 1H), 8.63 (d, J=5.26Hz, 1H), 7.96 (d, J=1.32Hz, 1H), 7.70 (s, 1H), 7.66 (s, 1H), 7.62 (dd, J=5.15, 1.43Hz, 1H), 5.66 (dd, J=5.92,3.07Hz, 1H), 4.37-4.50 (m, 2H), 4.04-4.14 (m, 1H), 3.66- 3.83 (m, 2H), 3.11-3.37 (m, 4H), 2.98-3.07 (m, 1H), 2.70 (s, 3H), 2.29-2.44 (m, 2H), 2.08 (dt, J=14.58,7.40Hz, 1H).

Embodiment 30

Synthesize (4S) -8- chloro- N- (2- ((S) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Under a nitrogen at 0 DEG C to the chloro- N- of (4S) -8- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) Methoxyl group) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diazaHCl/water solution is added in solution of -5 (the 2H)-formamides (6.5g, 11.00mmol) in methanol (65mL) (15mL, 494mmol, 36%), is then stirred at room temperature 2h.(TLC eluant, eluents：100% ethyl acetate:R_f=0.2；UV is activated ).Reactant mixture is alkalized (until pH-8-9) with saturated sodium bicarbonate solution at 0 DEG C and solvent is evaporated under reduced pressure.By remnants Thing is diluted with water (30mL) and is extracted into DCM (2x100mL).The organic extract of merging is through anhydrous sodium sulfate drying, filtering And evaporate filtrate, by rough thing with triturated under ether (50mL), obtain the chloro- N- of (4S) -8- (2- ((S) -2,3- the third oxygen of dihydroxy Base) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine- 5 (2H)-formamides (5.2g, 9.29mmol, 84% yield), it is pale solid.LCMS(m/z):551.06[M +H]⁺,R_t=2.14min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.05 (s, 1H), 8.32 (d, J=5.70Hz, 1H), 8.13-7.92 (m, 2H), 7.81-7.64 (m, 4H), 5.64 (dd, J=5.92,3.07Hz, 1H), 4.21-4.12 (m, 2H), 3.97 (dq, J= 9.95,5.09Hz, 1H), 3.73-3.57 (m, 2H), 3.38-3.10 (m, 4H), 3.02 (dd, J=12.28,3.29Hz, 1H), 2.50 (t, J=6.36Hz, 1H), 2.35 (dddd, J=14.17,10.00,5.97,4.06Hz, 1H), 2.14-2.00 (m, 1H)。

Embodiment 31

Synthesize (4S) -8- chloro- N- (4- ((R) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (4- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (3.2g, 5.42mmol) added in the solution of stirring in the methanol (30mL) hydrochloric acid (5mL, 165mmol), 5min time is lasted.Then reactant mixture is stirred into 2h at 30 DEG C.(TLC eluant, eluents：5%MeOH/DCM: R_f-0.5；UV activation).Solvent is evaporated and neutralized with sodium bicarbonate solution, the solid that obtains is filtered and with water and pentane (2x20mL) is washed, and obtains the chloro- N- of desired product (4S) -8- (4- ((R) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine (1.2g, 2.179mmol, 40.2% yield), it is white solid.LCMS(m/z):550.09[M+H]⁺, Rt= 1.94min。

¹H NMR(400MHz,CDCl₃):δ ppm 12.93 (s, 1H), 8.20 (s, 1H), 8.12 (d, J=7.67Hz, 1H), 8.04 (d, J=5.92Hz, 1H), 7.78-7.70 (m, 2H), 7.68-7.59 (m, 2H), 6.53 (dd, J=5.70,2.41Hz, 1H), 5.63 (dd, J=5.81,3.18Hz, 1H), 4.18-4.08 (m, 3H), 3.87-3.68 (m, 2H), 3.36-3.12 (m, 3H), 3.01 (dd, J=12.28,3.29Hz, 1H), 2.34 (dddd, J=14.11,10.00,6.03,4.06Hz, 1H), 2.15-2.01(m,2H),1.58(s,1H)。

Embodiment 32

Synthesize (4S) -8- chloro- N- (4- ((R) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-carboxamide hydrochlorides

At 0 DEG C to the chloro- N- of (4S) -8- (4- ((R) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (3- (fluoroforms Base) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (100mg, Ether HCl (2M) (4.55mL, 9.09mmol) 0.182mmol) is added in the solution of stirring in methanol (10mL), 5min is lasted Time.Then reactant mixture is stirred into 2h at 30 DEG C.(TLC eluant, eluents：10%MeOH/DCM:R_f-0.5；UV activation). Solvent is evaporated and washed (2 × 20mL) with pentane, the chloro- N- of desired product (4S) -8- (4- ((R) -2,3- dihydroxies are obtained Base propoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-carboxamide hydrochlorides (65mg, 0.110mmol, 60.5% yield), it is pale solid. LCMS(m/z):550.09[M+H],⁺R_t=1.94min.

¹H NMR(400MHz,DMSO-d₆):δppm 13.06(s,1H),8.22-8.07(m,4H),7.96-7.88(m, 1H), 7.79-7.85 (m, 1H), 7.18 (s, 1H), 6.91 (d, J=5.70Hz, 1H), 5.50 (dd, J=5.92,3.07Hz, 1H), 4.13 (dd, J=9.98,3.84Hz, 1H), 3.97 (dd, J=9.87,6.36Hz, 1H), 3.90-3.80 (m, 3H), 3.53-3.23 (m, 6H), 2.43-2.28 (m, 1H), 2.11 (dt, J=13.65,7.10Hz, 1H).

Embodiment 33

Synthesize (4S) -8- chloro- N- (5- ((S) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to the chloro- N- of (4S) -8- (5- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaHCl/water solution is added in the solution of stirring of -5 (the 2H)-formamides (500mg, 0.846mmol) in methanol (10mL) (2mL, 65.8mmol) and stir 1h.(TLC systems:5%MeOH/DCM, Rf value：0.3).Reactant mixture is evaporated to remove Methanol, then at 10 DEG C by residue saturation NaHCO₃(10mL) is basified, and the solid of acquisition is filtered, is washed with water (10mL) and in high vacuum dry, the desired pure chloro- N- of product (4S) -8- (5- ((S) -2,3- dihydroxy propoxyl group) are obtained Pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (300mg, 0.543mmol, 64.2% yield), it is light yellow solid.LCMS(m/z):551.03[M +H]⁺, Rt=2.35min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.95 (s, 1H), 8.89 (d, J=1.32Hz, 1H), 8.14 (s, 1H), 8.06 (d, J=8.11Hz, 1H), 7.93 (d, J=1.32Hz, 1H), 7.57-7.78 (m, 3H), 5.67 (dd, J=5.92, 3.07Hz, 1H), 4.32-4.52 (m, 2H), 4.07 (dq, J=9.65,4.75Hz, 1H), 3.64-3.80 (m, 2H), 3.11- 3.37 (m, 4H), 3.02 (dd, J=12.28,3.29Hz, 1H), 2.22-2.47 (m, 2H), 2.09 (dt, J=14.20, 7.04Hz,1H)。

Embodiment 34

Synthesize (4S) -8- chloro- N- (5- ((R) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to the chloro- N- of (4S) -8- (5- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaHCl/water solution is added in the solution of stirring of -5 (the 2H)-formamides (500mg, 0.846mmol) in methanol (10mL) (0.026mL, 0.846mmol) and stir 1h.(TLC eluant, eluents：5%MeOH/DCM Rf:0.3；UV activation).Reaction is mixed Compound is evaporated to remove methanol, and residue uses saturation NaHCO at 10 DEG C₃Alkalized (10mL) solution, and the solid of acquisition is filtered, and is used Water washing (10mL) and in high vacuum dry, obtains the desired pure chloro- N- of product (4S) -8- (5- ((R) -2,3- dihydroxy third Epoxide) pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] Diaza- 5 (2H)-formamides (350mg, 0.631mmol, 74.6% yield), it is light yellow solid.LCMS(m/z): 551.03[M+H]⁺；Rt=2.45min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.95 (s, 1H), 8.89 (d, J=1.32Hz, 1H), 8.14 (s, 1H), 8.06 (d, J=7.67Hz, 1H), 7.93 (d, J=1.32Hz, 1H), 7.56-7.79 (m, 3H), 5.67 (dd, J=5.92, 3.07Hz, 1H), 4.34-4.53 (m, 2H), 4.07 (dq, J=9.87,4.97Hz, 1H), 3.64-3.82 (m, 2H), 3.11- 3.36 (m, 4H), 3.02 (dd, J=12.28,3.29Hz, 1H), 2.23-2.44 (m, 2H), 1.94-2.20 (m, 1H).

Embodiment 35

Synthesize (4S) -8- chloro- N- (pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diazaNaH is added in the solution of the stirring of (300mg, 0.883mmol) in THF (20mL) (31.8mg,1.325mmol).Then reactant mixture is stirred into 30min at 30 DEG C and adds 3- (pyridine -2- bases) -2H- pyridines And [1,2-a] [1,3,5] triazine -2,4 (3H)-diketone (255mg, 1.060mmol).Reactant mixture is stirred into 15h at 70 DEG C. (TLC:10%MeOH/ ethyl acetate, Rf:0.8).Room temperature is allowed to cool to, and (20mL) is diluted with cold water, ethyl acetate is used Extract (2X40mL).The organic layer of merging is through anhydrous Na₂SO₄Dry and be concentrated in vacuo, obtain crude compound.Crude mixture Through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：90%EtOAc/ petroleum ethers), then carry out preparation HPLC (MP-A:5mM ammonium hydrogen carbonate (aqueous solution) MP-B：Acetonitrile post：The μ methods of symmetryc8 (300x21.2) 7：T/%B:ISO(20: 80) flow velocity：18ml/min eluant, eluents：Excessive THF+ACN+MEOH), obtain the chloro- N- of desired product (4S) -8- (pyridine -2- Base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (205mg, 0.446mmol, 50.5% yield), it is pale solid.LCMS(m/z):460.01[M+H ]⁺,Rt:2.83min。

¹H NMR(400MHz,CDCl₃):δppm 12.91(s,1H),8.26-8.20(m,2H),8.16-8.06(m,2H), 7.73 (d, J=7.67Hz, 1H), 7.68-7.58 (m, 3H), 6.95 (ddd, J=7.29,4.88,0.99Hz, 1H), 5.68 (dd, J=6.03,3.18Hz, 1H), 3.37-3.13 (m, 3H), 3.06-2.94 (m, 1H), 2.34 (dddd, J=14.20, 10.08,6.08,4.06Hz,1H),2.17-1.98(m,1H)。

Embodiment 36

Synthesize (4S) -8- methyl -7- (2- picoline -4- bases)-N- (pyridine -2- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -8- methyl -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diazaNaH is added in the solution of (300mg, 1.126mmol) in tetrahydrofuran (THF) (15mL) (54.1mg,2.253mmol).Then reactant mixture is stirred into 1h at 30 DEG C under a nitrogen.Add 3- (pyridine -2- bases) -2H- Pyrido [1,2-a] [1,3,5] triazine -2,4 (3H)-diketone (541mg, 2.253mmol) then by reactant mixture at 70 DEG C Stir 15h.(TLC systems；.Rf-0.5,5% ethanol/methylene).So that reactant mixture is cooled to room temperature and pours into cold water In (30mL), it is extracted with ethyl acetate (2x50mL).The organic layer of merging obtains thick through anhydrous sodium sulfate drying and vacuum concentration Product.Crude mixture is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：3%MeOH/EtOAc) and further (condition is purified by preparation HPLC：MP-A:5mM ammonium hydrogen carbonate (aqueous solution) MP-B：Acetonitrile post：Xterra C18 (250x19) 10u methods：50:50 flow velocitys：18ml/min eluant, eluents：ACN+THF), obtain desired product (4S) -8- methyl - 7- (2- picoline -4- bases)-N- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (132mg, 0.339mmol, 30.1% yield), it is pale solid.LCMS(m/z):387.0 [M+H]^+,Rt:3.41min。

¹H NMR(400MHz,CDCl₃):δ ppm 13.28 (s, 1H), 8.61 (d, J=5.26Hz, 1H), 8.31-8.24 (m, 1H), 8.13 (d, J=8.55Hz, 1H), 7.71-7.62 (m, 2H), 7.45 (s, 1H), 7.39 (dd, J=5.04, 1.53Hz, 1H), 6.95 (ddd, J=7.29,4.99,0.88Hz, 1H), 5.66 (dd, J=5.81,3.18Hz, 1H), 3.34- 3.11 (m, 3H), 3.05-2.96 (m, 1H), 2.71 (s, 3H), 2.42 (s, 3H), 2.36-2.26 (m, 1H), 2.06 (dt, J= 14.03,7.02Hz,1H)。

Embodiment 37

Synthesize (4S) -8- chloro- N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridin-4-yl) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaIn the solution of stirring of -5 (the 2H)-formamides (350mg, 0.652mmol) in methanol (5mL) be added dropwise HCl (3.96 μ L, 0.130mmol), 5min time is lasted.Then reactant mixture is stirred into 2h at 30 DEG C.(TLC eluant, eluents：5%MeOH/ DCM:R_f- 0.6) and it is concentrated in vacuo, uses saturation NaHCO₃Solution is neutralized, and filters the solid obtained, (2x20mL) is washed with ether, Obtain the chloro- N- of desired product (4S) -8- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (2- picolines - 4- yls) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (180mg, 0.361mmol, 55.3% yield), it is pale solid.LCMS(m/z):497.10[M+H]⁺,Rt:1.41min。

¹H NMR(400MHz,DMSO-d₆):δ ppm 12.71 (s, 1H), 8.66 (d, J=5.04Hz, 1H), 7.98 (d, J =5.70Hz, 1H), 7.88 (s, 1H), 7.62 (s, 1H), 7.58 (d, J=5.26Hz, 1H), 6.85 (d, J=1.53Hz, 1H), 6.80-6.76 (m, 1H), 5.44 (dd, J=5.92,2.85Hz, 1H), 4.85 (d, J=5.04Hz, 1H), 4.59 (t, J= 5.70Hz, 1H), 4.22 (dd, J=10.85,4.49Hz, 1H), 4.11 (dd, J=10.96,6.14Hz, 1H), 3.76 (dq, J =10.61,5.45Hz, 1H), 3.42 (t, J=5.81Hz, 2H), 3.23-3.29 (m, 1H), 3.14-3.06 (m, 2H), 3.02- 2.94(m,1H),2.61(s,3H),2.29-2.19(m,1H),2.01-1.90(m,1H)。

Embodiment 38

Synthesize (4S) -8- chloro- N- (4- ((R) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (4- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaIn the solution of stirring of -5 (the 2H)-formamides (180mg, 0.335mmol) in methanol (8mL) be added dropwise HCl (0.102mL, 3.35mmol), 5min time is lasted.Then reactant mixture is stirred into 2h at 30 DEG C.(TLC eluant, eluents：5%MeOH/DCM: R_f-0.1；UV activation), and evaporation solvent.Reactant mixture is neutralized and filtered with sodium bicarbonate solution the solid obtained, is used Ether washs (2x15mL), obtains the chloro- N- of desired product (4S) -8- (4- ((R) -2,3- dihydroxy propoxyl group) pyridine -2- Base) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (88mg, 0.174mmol, 51.8% yield), it is brown solid.LCMS(m/z):497.1[M+H]⁺,Rt =1.26min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.91 (s, 1H), 8.62 (d, J=4.82Hz, 1H), 8.08 (d, J= 5.48Hz, 1H), 7.78 (d, J=18.20Hz, 2H), 7.69-7.62 (m, 2H), 6.56 (d, J=3.73Hz, 1H), 4.13 (s, 3H), 3.89-3.70 (m, 2H), 3.37-3.10 (m, 3H), 3.01 (d, J=12.50Hz, 1H), 2.71 (m, 4H), 2.65- 2.62(m,1H),2.33(s,1H),2.21(s,1H),2.14-2.00(m,1H)。

Embodiment 39

Synthesize (4S) -8- chloro- N- (2- ((R) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides:

At 0 DEG C to the chloro- N- of (4S) -8- (2- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridin-4-yl) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaHCl/water solution is added in the solution of stirring of -5 (the 2H)-formamides (350.0mg, 0.652mmol) in methanol (5.0mL) (0.990mL, 32.6mmol) and stirring 5h.Reactant mixture is alkalized (until pH-8- with saturated sodium bicarbonate solution at 0 DEG C 9) and concentrate.By residue diluted with water (8mL) and it is extracted into EtOAc (3x10mL).The organic extract of merging is through anhydrous Sodium sulphate is dried, and is filtered and is evaporated filter vacuum, obtains the pure chloro- N- of (4S) -8- (2- ((R) -2,3- dihydroxy propoxyl group) Pyridin-4-yl) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (190.5mg, 0.375mmol, 57.5% yield), it is pale solid.LCMS(m/z):497.10 [M+H]⁺,R_t=1.40min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.80 (s, 1H), 8.70 (d, J=5.70Hz, 1H), 7.90 (d, J= 5.92Hz, 1H), 7.68 (s, 1H), 7.49-7.45 (m, 2H), 6.94 (d, J=1.75Hz, 1H), 6.85 (dd, J=5.92, 1.75Hz, 1H), 5.64 (dd, J=6.03,3.18Hz, 1H), 4.43-4.41 (m, 2H), 4.29 (d, J=5.48Hz, 1H), 4.01-3.95(m,1H),3.72-3.60(m,2H),3.35-3.19(m,2H),3.16-3.10(m,1H),3.05-3.00(m, 1H), 2.92 (t, J=6.47Hz, 1H), 2.69 (s, 3H), 2.40-2.30 (m, 1H), 2.07 (dt, J=14.20,7.26Hz, 1H)。

Embodiment 40

Synthesize (4S) -8- chloro- N- (5- ((S) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to the chloro- N- of (4S) -8- (5- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaAdded in the solution of stirring of -5 (the 2H)-formamides (450mg, 0.836mmol) in methanol (20mL) 2.0M HCl (5mL, 10.00mmol).1h is stirred at room temperature in gained reactant mixture.(TLC systems:10%MeOH/DCM, Rf:0.4).Will reaction Mixture is concentrated under reduced pressure to remove methanol, is then alkalized with saturated sodium bicarbonate solution (20mL), is extracted with DCM (3X30mL). The organic layer of merging is with water (20mL), saline solution (20mL) washing and through anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain The semisolid obtained is washed with ether, is obtained solid chemical compound, is filtered, obtain the chloro- N- (5- of desired product (4S) -8- ((S) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (205mg, 0.407mmol, 48.7% yield), it is faint yellow Solid.LCMS(m/z):498.14[M+H]⁺,R_t=1.14min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.95 (s, 1H) 8.92 (d, J=1.32Hz, 1H) 8.63 (d, J= 5.26Hz, 1H) 7.97 (d, J=1.32Hz, 1H) 7.71 (s, 1H) 7.67 (s, 1H), 7.63 (d, J=5.04Hz, 1H) 5.66 (dd, J=5.81,3.18Hz, 1H) 4.50-4.36 (m, 2H) 4.09 (br s, 1H) 3.81-3.66 (m, 2H) 3.37-3.11 (m, 4H) 3.06-2.99 (m, 1H) 2.69 (s, 3H) 2.46-2.31 (m, 2H) 2.09 (dt, J=14.03,7.02Hz 1H).

Embodiment 41

Synthesize (4S) -8- chloro- N- (6- ((R) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (6- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaHCl/water solution (6mL, 12.00mmol, 2M) is added in the solution of stirring of -5 (the 2H)-formamides in methanol (6mL), is gone through When 10min a period of time, 30min is stirred at room temperature in settled solution.(TLC systems:10% methanol/DCM.R_f:0.2).Will Reactant mixture is concentrated in vacuo, and is obtained clear yellow viscous oily thing and is neutralized with saturated sodium bicarbonate solution, obtains orange precipitation.Will Orange solids are filtered and are dried in vacuo, and obtain the chloro- N- of (4S) -8- (6- ((R) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine (159mg, 0.316mmol, 56.6% yield), it is orange solids.LCMS(m/z):497.1[M+H]⁺, Rt= 1.54min。

¹H NMR(400MHz,DMSO-d₆):δ ppm 12.44 (s, 1H), 8.65 (d, J=5.04Hz, 1H), 7.85 (s, 1H), 7.57-7.71 (m, 2H), 7.40-7.57 (m, 2H), 6.44 (dd, J=7.67,1.10Hz, 1H), 5.48 (dd, J= 5.81,3.18Hz, 1H), 4.76 (br s, 1H), 4.57 (br s, 1H), 3.76 (br d, J=7.23Hz, 1H), 3.61-3.71 (m, 2H), 3.32-3.49 (m, 2H), 3.22 (m, 1H), 3.04-3.19 (m, 2H), 2.97 (br dd, J=12.06,3.07Hz, 1H),2.53-2.58(m,3H),2.22(m,1H),1.84-1.95(m,1H)。

Embodiment 42

Synthesize (4S) -8- chloro- N- (2- (3- hydroxy propyloxy groups) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diazaThree are added in the solution of the stirring of (300mg, 0.883mmol) in tetrahydrofuran (10mL) Phosgene (157mg, 0.530mmol) and triethylamine (0.738mL, 5.30mmol) and stirring 30min.Then at 28 DEG C by 3- ((4- Aminopyrimidine -2- bases) epoxide) propyl- 1- alcohol (299mg, 1.766mmol) adds to reactant mixture, then stirs 16h at 80 DEG C. (TLC systems:5% methanol/ethyl acetate .R_fValue：0.4,UV).So that reactant mixture is cooled to room temperature and is diluted with water (40mL), is extracted with ethyl acetate (2X50mL).The organic layer of merging is dry through anhydrous sodium sulfate with aqueous salt solu-tion (30mL) It is dry, filter and concentrate, obtain crude compound.Crude compound is through flash column chromatography (silica gel:100-200 mesh, elution Agent：80% ethyl acetate/petroleum ether), obtain the chloro- N- of desired product (4S) -8- (2- (3- hydroxy propyloxy groups) pyrimidine -4- Base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (65mg, 0.117mmol, 13.27% yield), it is pale solid.LCMS(m/z):535.15[M+H ]⁺, Rt=2.38min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 12.97 (s, 1H), 8.40 (d, J=5.70Hz, 1H), 8.07 (d, J =7.89Hz, 1H), 8.04 (s, 1H), 7.92-7.86 (m, 2H), 7.83-7.78 (m, 1H), 7.64 (d, J=5.70Hz, 1H), 5.46 (dd, J=5.70,3.07Hz, 1H), 4.48 (t, J=5.15Hz, 1H), 3.97-3.87 (m, 2H), 3.50-3.42 (m, 2H), 3.25 (s, 1H), 3.17-3.06 (m, 2H), 2.99 (dd, J=12.06,3.29Hz, 1H), 2.34-2.19 (m, 1H), 2.04-1.92 (m, 1H), 1.72 (quin, J=6.30Hz, 2H).

Embodiment 43

Synthesize (4S) -8- chloro- N- (6- (3- hydroxyl -2,2- dimethyl propylenes epoxide) pyrazine -2- bases) -7- (3- (fluoroforms Base) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 10 DEG C to 3- ((6- Aminopyrazine -2- bases) epoxide) -2,2- dimethyl propylene -1- alcohol (406mg, 2.060mmol) In the solution of stirring in tetrahydrofuran (15mL) add triphosgene (183mg, 0.618mmol) and triethylamine (0.862mL, 6.18mmol) and stirring 30min.Then 28 DEG C by the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysenes - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(350mg, 1.030mmol) adds to reactant mixture, Ran Hou 80 DEG C of stirring 16h.(TLC systems:5% methanol/ethyl acetate .R_fValue：0.4,UV).Make reactant mixture be cooled to room temperature to be used in combination Water dilutes (40mL), is extracted with ethyl acetate (2X70mL).The organic layer of merging is with aqueous salt solu-tion (30mL), through anhydrous sulphur Sour sodium is dried, filtered and concentrated, and obtains crude compound.Crude compound is through flash column chromatography (silica gel:100-200 mesh, 100% ethyl acetate), obtain the chloro- N- of desired product (4S) -8- (6- (3- hydroxyl -2,2- dimethyl propylenes epoxide) pyrazine -2- Base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (146mg, 0.256mmol, 24.83% yield), it is pale solid.LCMS(m/z):563.19[M+H ]⁺,R_t=2.71min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.67 (s, 1H), 8.88 (s, 1H), 8.03 (s, 1H), 7.96 (d, J= 7.67Hz, 1H), 7.90 (s, 1H), 7.75-7.70 (m, 1H), 7.70-7.61 (m, 2H), 5.70 (dd, J=5.92,3.07Hz, 1H), 3.74-3.63 (m, 2H), 3.36-3.20 (m, 4H), 3.11-3.19 (m, 2H), 3.02 (dd, J=12.17,3.18Hz, 1H), 2.43-2.30 (m, 1H), 2.09 (dt, J=14.31,7.21Hz, 1H), 0.87 (d, J=1.75Hz, 6H).

Embodiment 44

Synthesize (4S) -8- chloro- N- (6- ((S) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyrimidine-4-yl) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaIn the solution of stirring of -5 (the 2H)-formamides (185mg, 0.344mmol) in methanol (5mL) add hydrochloric acid (0.145mL, 1.719mmol), 2h is then stirred at room temperature.(TLC eluant, eluents：10%MeOH/DCM Rf:0.4；UV activation).Reaction is mixed Compound is concentrated in vacuo and by residue NaHCO₃The aqueous solution neutralizes and filters the solid of acquisition, is then washed with pentane (2x10mL), obtains the chloro- N- of desired product (4S) -8- (6- ((S) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (2- Picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (148mg, 0.284mmol, 82% yield), it is Light brown solid.LCMS(m/z):498.0[M+H]⁺, Rt=1.38min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.03 (s, 1H), 8.62 (d, J=5.26Hz, 1H), 8.47 (d, J =0.88Hz, 1H), 7.88 (s, 1H), 7.70 (s, 1H), 7.65 (dd, J=5.26,1.32Hz, 1H), 7.41 (d, J= 0.88Hz, 1H), 5.45 (dd, J=5.92,3.07Hz, 1H), 4.97 (s, 1H), 4.67 (br s, 1H), 4.35 (dd, J= 10.74,4.17Hz, 1H), 4.20 (dd, J=10.85,6.47Hz, 1H), 3.82-3.77 (m, 1H), 3.46-3.41 (m, 2H), 3.27-3.23 (m, 1H), 3.16-3.09 (m, 2H), 2.98 (dd, J=12.06,3.29Hz, 1H), 2.61 (s, 3H), 2.25 (td, J=6.85,4.06Hz, 1H), 2.01-1.93 (m, 1H).

Embodiment 45

Synthesize (4S) -8- chloro- N- (6- ((S) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to the chloro- N- of (4S) -8- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaAdd 4M HCl's in the solution of stirring of -5 (the 2H)-formamides (6.0g, 10.15mmol) in 1,4- dioxanes (30mL) Dioxane solution (15.06g, 50.8mmol) and it is stirred for 4h.(TLC systems:100% ethyl acetate .Rf values：0.2).Will reaction Mixture evaporates, and adds saturation NaHCO₃Solution (50mL), filters the solid obtained and vacuum drying, obtains desired product The chloro- N- of (4S) -8- (6- ((S) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- two Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(5.4g, 9.80mmol, 97% are received -5 (2H)-formamides Rate), it is white solid.LCMS(m/z):551.13[M+H]⁺,R_t=2.18min.

¹H NMR(400MHz,DMSO-d₆):δppm 12.69(s,1H),8.87(s,1H),8.05-7.99(m,2H), 7.94-7.80 (m, 4H), 5.50 (dd, J=5.92,3.07Hz, 1H), 4.89 (d, J=5.04Hz, 1H), 4.62 (t, J= 5.59Hz,1H),3.83-3.66(m,3H),3.40-3.23(m,3H),3.18-3.07(m,2H),3.01-2.94(m,1H), 2.30-2.20 (m, 1H), 1.98 (dt, J=14.14,7.18Hz, 1H).

Embodiment 46

Synthesize (4S) -8- chloro- N- (4- ((R) -2,3- dihydroxy propoxyl group) pyrimidine -2-base) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to the chloro- N- of (4S) -8- (4- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyrimidine -2-base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaHCl/water solution is added in the solution of stirring of -5 (the 2H)-formamides (200mg, 0.338mmol) in methanol (5mL) (0.294mL, 3.38mmol) and stirring 2h.(TLC systems:100% ethyl acetate, Rf values：0.3).Then by reactant mixture Use saturation NaHCO₃Solution is quenched (10mL) and extracted (2x30mL) with DCM.The organic layer of merging is through anhydrous Na₂SO₄Dry, mistake Filter simultaneously filtrate is evaporated, obtain the chloro- N- of desired product (4S) -8- (4- ((R) -2,3- dihydroxy propoxyl group) pyrimidine -2-base) - 7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first Acid amides (130mg, 0.236mmol, 69.6% yield), it is white solid.LCMS(m/z):551.10[M+H]⁺, Rt= 1.92min。

¹H NMR(400MHz,DMSO-d₆):δ ppm 12.94 (s, 1H), 8.22 (d, J=5.70Hz, 1H), 8.11-8.05 (m, 2H), 7.88-7.75 (m, 3H), 6.55 (d, J=5.70Hz, 1H), 5.46 (dd, J=6.03,2.96Hz, 1H), 4.93 (d, J=4.82Hz, 1H), 4.63 (t, J=5.70Hz, 1H), 4.23-4.16 (m, 1H), 4.13-4.08 (m, 1H), 3.78- 3.71 (m, 1H), 3.42-3.34 (m, 2H), 3.28-3.24 (m, 1H), 3.16-3.06 (m, 2H), 2.98 (dd, J=12.06, 3.29Hz, 1H), 2.29-2.19 (m, 1H), 1.95 (dt, J=14.09,7.32Hz, 1H).

Embodiment 47

Synthesize (4S) -8- chloro- N- (3- fluoro- 5- (pyridin-3-yl) phenyl) -7- (2- picoline -4- bases) -3,4- two Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In seal pipe, in room temperature to the chloro- 7- of (4S) -8- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-bridge Methylene pyridine simultaneously [2,3-b] [1,4] diazaIn the solution of the stirring of (300mg, 1.046mmol) in THF (15mL) Add triphosgene (186mg, 0.628mmol) and triethylamine (0.875mL, 6.28mmol) and stirring 30min.Then, 3- is added Fluoro- 5- (pyridin-3-yl) aniline (236mg, 1.255mmol), then stirs 15h at 80 DEG C.(TLC systems:Rf-0.3,5% MeOH/EtOAc).So that reactant mixture is cooled to room temperature and is diluted with water (25mL), extracted with EtOAc (2x40mL).Merge Organic layer through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain crude compound.Crude compound is through quick post color Spectrum purifying (silica gel：100-200 mesh, eluant, eluent：1% methanol/ethyl acetate), obtain the chloro- N- (3- of desired product (4S) -8- Fluoro- 5- (pyridin-3-yl) phenyl) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (150mg, 0.295mmol, 28.2% yield), it is yellow solid.LCMS(m/ z):501.09[M+H]⁺,R_t=1.89min.

¹H NMR(400MHz,CDCl3):δppm 12.80(s,1H),8.77-8.74(m,1H),8.68-8.65(m, 1H), 8.63 (dd, J=4.82,1.75Hz, 1H), 7.71-7.66 (m, 2H), 7.50-7.45 (m, 2H), 7.40-7.30 (m, 3H), 7.00-6.95 (m, 1H), 5.67 (dd, J=5.81,3.18Hz, 1H), 3.37-3.12 (m, 3H), 3.06-3.00 (m, 1H),2.55(s,3H),2.40-2.31(m,1H),2.14-2.05(m,1H)。

Embodiment 48

Synthesize (4S) -8- chloro- N- (2- (3- hydroxy-3-methyls butoxy) pyrimidine-4-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In seal pipe, in room temperature to the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysenes-Isosorbide-5-Nitrae-bridge Methylene pyridine simultaneously [2,3-b] [1,4] diazaIn the solution of the stirring of (300mg, 0.883mmol) in THF (15mL) Add triphosgene (157mg, 0.530mmol) and triethylamine (0.738mL, 5.30mmol) and stirring 30min.Then, 4- is added ((4- aminopyrimidine -2- bases) epoxide) -2- methyl butyl- 2- alcohol (348mg, 1.766mmol), then stirs 15h at 80 DEG C.(TLC System:R_f- 0.4,5%MeOH/EtOAc).So that reactant mixture is cooled to room temperature and is diluted with water (25mL), extracted with EtOAc Take (2x40mL).The organic layer of merging is through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain crude compound.Roughization Compound is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：1% methanol/ethyl acetate), obtain desired product The chloro- N- of (4S) -8- (2- (3- hydroxy-3-methyls butoxy) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamide (110mg, 0.192mmol, 21.75% Yield), it is pale solid.LCMS(m/z):563.12[M+H]⁺,R_t=2.58min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.01 (s, 1H), 8.34 (d, J=5.70Hz, 1H), 8.07-7.97 (m, 2H), 7.76-7.63 (m, 4H), 5.64 (dd, J=5.92,3.07Hz, 1H), 4.30-4.11 (m, 2H), 3.43-3.11 (m, 3H), 3.02 (dd, J=12.28,3.29Hz, 1H), 2.40-2.28 (m, 1H), 2.13-2.04 (m, 2H), 1.88 (t, J= 6.25Hz,2H),1.26(s,6H)。

Embodiment 49

Synthesize (4S) -8- chloro- N- (6- (3- hydroxyl -2,2- dimethyl propylenes epoxide) pyridine -2- bases) -7- (3- (fluoroforms Base) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Under a nitrogen to the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diazaThree second are added in the solution of the stirring of (300mg, 0.883mmol) in THF (15mL) Amine (0.738mL, 5.30mmol), triphosgene (262mg, 0.883mmol), is then stirred at room temperature 30min.Mixed to the reaction 3- ((6- aminopyridine -2- bases) epoxide) -2,2- dimethyl propylene -1- alcohol (520mg, 2.65mmol) is added in thing, then 70 DEG C heating 16h.(TLC eluant, eluents：100%EtOAc/ hexanes, Rf:0.3, UV activation).Reactant mixture is cooled to RT, plus Enter water (10mL) and extracted (3x10mL) with EtOAc.The organic extract of merging is filtered and evaporated through anhydrous sodium sulfate drying To crude product.Thick material is purified into (condition-post by preparation HPLC：Xterra(250X21.1mm,10μ)；Mobile phase-A: 10mM ammonium hydrogen carbonate；Mobile phase-B:Acetonitrile；Column temperature：Environment temperature；Flow velocity:18ml/min:Diluent：THF+MEOH+ACN), Obtain the chloro- N- of (4S) -8- (6- (3- hydroxyl -2,2- dimethyl propylenes epoxide) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (128mg, 0.224mmol, 25.3% yield).LCMS(m/z):562.11[M+H]⁺,R_t=2.93min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.56 (s, 1H), 8.06 (s, 1H), 8.01 (d, J=7.67Hz, 1H), 7.73-7.61 (m, 3H), 7.57-7.51 (m, 2H), 6.45-6.38 (m, 1H), 5.68 (dd, J=5.92,3.07Hz, 1H), 3.67 (q, J=11.18Hz, 2H), 3.35-3.18 (m, 2H), 3.13 (br d, J=7.02Hz, 3H), 3.00 (dd, J= 12.28,3.29Hz, 1H), 2.79 (br t, J=6.91Hz, 1H), 2.39-2.27 (m, 1H), 2.14-2.03 (m, 1H), 0.79 (d, J=5.04Hz, 6H).

Embodiment 50

Synthesize (4S) -8- chloro- N- (2- ((R) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamide

At 0 DEG C to the chloro- N- of (4S) -8- (2- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) first under nitrogen Epoxide) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] DiazaHCl/water solution is added in the solution of stirring of -5 (the 2H)-formamides (20g, 33.8mmol) in methanol (100mL) (10mL, 40.0mmol, 36%), is then stirred at room temperature 1h.(TLC eluant, eluents：5% methanol/DCM, Rf:0.3, UV activation ).Reactant mixture is concentrated, residue saturation NaHCO₃Basified (until pH-8-9) and 15min is stirred at room temperature. Gained solid by filtration is separated and absorbed in DMSO (20mL) and water (600mL) mixture, and 16h is then stirred at room temperature. Solid is filtered, dries, obtains the pure chloro- N- of (4S) -8- (2- ((R) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (15.3g, 27.7mmol, 82% yield), it is pale solid.LCMS(m/z):551.13[M+H]⁺,R_t=2.15min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.02 (s, 1H), 8.30 (d, J=5.70Hz, 1H), 8.06-7.97 (m, 2H), 7.80-7.67 (m, 4H), 5.62 (dd, J=5.92,3.07Hz, 1H), 4.18-4.04 (m, 2H), 3.97 (dq, J= 9.92,5.10Hz, 1H), 3.77 (d, J=5.48Hz, 1H), 3.71-3.58 (m, 2H), 3.37-3.20 (m, 2H), 3.19- 3.10 (m, 2H), 3.02 (dd, J=12.28,3.29Hz, 1H), 2.40-2.29 (m, 1H), 2.14-2.01 (m, 1H).

Embodiment 51

Synthesize (4S) -8- chloro- N- (2- ((R) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-carboxamide hydrochlorides

Under a nitrogen at 0 DEG C to the chloro- N- of (4S) -8- (2- ((R) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides Added in the solution of the stirring of (250mg, 0.454mmol) in ether (5mL) 2.0M HCl diethyl ether solution (3mL, 6.00mmol), 1h is then stirred at room temperature.(TLC eluant, eluents：5%MeOH/DCM, R_f- 0.3, UV activation).By solvent under reduced pressure Evaporation, by rough thing with triturated under ether (2x10mL), obtaining the chloro- N- of (4S) -8-, (2- ((R) -2,3- dihydroxy propoxyl group) is phonetic Pyridine -4- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-carboxamide hydrochlorides (220mg, 0.369mmol, 81% yield), it is faint yellow solid.LCMS(m/z):551.0 [M+H]⁺,R_t=3.45min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 12.93 (s, 1H), 8.45 (d, J=5.70Hz, 1H), 8.15-7.97 (m, 3H), 7.93-7.77 (m, 2H), 7.69 (d, J=5.70Hz, 1H), 5.80 (brs, 4H), 5.50 (br dd, J=5.48, 2.85Hz,1H),4.03-3.82(m,2H),3.77-3.64(m,1H),3.54-3.09(m,6H),2.42-2.28(m,1H), 2.07 (dt, J=13.92,7.07Hz, 1H).

Embodiment 52

Synthesize (4S) -8- chloro- N- cyclopropyl -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamide

In room temperature to the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diazaAdd in the solution of the stirring of (15g, 44.2mmol) in THF (600mL, in seal pipe) Enter triphosgene (6.55g, 22.08mmol) and DIPEA (38.6mL, 221mmol) and stirring 30min.Then cyclopropylamine is added (6.30mL, 88mmol), then stirs 16h at 80 DEG C.(TLC systems:Rf-0.4,90%EtOAc- petroleum ethers).So that reaction Mixture is cooled to room temperature, and (500ml) is diluted with water, and is extracted with ethyl acetate (3x900ml).The organic layer of merging is through nothing Aqueous sodium persulfate is dried and is concentrated under reduced pressure, and obtains crude compound.Crude product is through flash column chromatography (silica gel：100-200 mesh, Eluant, eluent：80% ethyl acetate/hexane), obtain desired product (4S) -8- chloro- N- cyclopropyl -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (5.1g, 11.96mmol, 27.1% yield), it is pale solid.LCMS(m/z):423.0[M+H]⁺, Rt=2.65min.

¹H NMR(400MHz,CDCl₃):δ ppm 10.05 (br s, 1H), 7.94-7.85 (m, 2H), 7.71 (d, J= 7.89Hz, 1H), 7.63-7.53 (m, 2H), 5.63 (dd, J=5.92,3.07Hz, 1H), 3.34-3.13 (m, 2H), 3.11- 3.02 (m, 1H), 2.97-2.90 (m, 1H), 2.77 (tq, J=7.13,3.65Hz, 1H), 2.27 (dddd, J=14.09, 10.03,6.14,3.95Hz,1H),2.05-1.96(m,1H),0.78-0.65(m,2H),0.46-0.35(m,2H)。

Embodiment 53

Synthesize (4S) -8- chloro- N- cyclopropyl -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-carboxamide hydrochlorides

At 0 DEG C to the chloro- N- cyclopropyl -7- of (4S) -8- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (100mg, 0.237mmol) are in 1,4- dioxanes (3mL) Solution and stirring 2h of the 4M HCl in 1,4 dioxanes (0.473mL, 1.892mmol) are added in the solution of stirring.(TLC systems System:Rf-0.1, EtOAc) in room temperature.Reactant mixture is concentrated under reduced pressure, residue is obtained, and it is ground with pentane (3 × 10mL).Gained solid is filtered through Buchner funnel, and (5mL) is rinsed with pentane, obtains the chloro- N- rings of desired product (4S) -8- Propyl group -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-carboxamide hydrochloride (50mg, 0.108mmol, 45.7% yield), it is pale solid.LCMS(m/z):423.0[M +H]⁺, Rt=2.67min.

¹H NMR(400MHz,CDCl₃):δppm 9.65(s,1H),8.57(s,1H),7.96-7.86(m,2H),7.79 (d, J=7.89Hz, 1H), 7.70-7.63 (m, 1H), 5.91 (s, 1H), 4.03 (s, 1H), 3.81-3.69 (m, 2H), 3.59 (s, 1H), 2.82-2.62 (m, 2H), 2.47 (s, 1H), 0.77 (q, J=6.36Hz, 2H), 0.49-0.39 (m, 2H).

Embodiment 54

Synthesize (4S) -8- chloro- N- (6- ((S) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-carboxamide hydrochlorides

At 0 DEG C to the chloro- N- of (4S) -8- (6- ((S) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (3- (fluoroforms Base) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (50mg, HCl (0.028mL, 0.908mmol) 0.091mmol) is added in the solution of the stirring in methanol (5mL), is then stirred in room temperature Mix 4h.(TLC systems:Rf-0.1,EtOAc).Reactant mixture is concentrated under reduced pressure, residue is obtained.By residue pentane Grind (3 × 10mL).Gained solid is filtered through Buchner funnel, and (5mL) is rinsed with pentane, obtain desired product (4S)- The chloro- N- of 8- (6- ((S) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza(20mg, 0.034mmol, 37.4% are received -5 (2H)-carboxamide hydrochlorides Rate), it is pale solid.LCMS(m/z):551.1[M+H]⁺,R_t=2.19min.

¹H NMR(400MHz,DMSO-d₆):δppm 12.62(s,1H),8.86(s,1H),8.05-7.99(m,3H), 7.94 (s, 1H), 7.89-7.84 (m, 2H), 5.54 (dd, J=5.70,2.85Hz, 1H), 3.82-3.67 (m, 5H), 3.44- 3.14(m,6H),2.38-2.28(m,1H),2.12-2.02(m,1H)。

Embodiment 55

Synthesize (4S) -8- chloro- N- (2- ((tetrahydrochysene -2H- pyrans -4- bases) epoxide) pyrimidine-4-yl) -7- (3- (fluoroforms Base) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamide

In seal pipe, in room temperature to the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diazaIn the solution of the stirring of (300mg, 0.883mmol) in THF (10mL) Triphosgene (131mg, 0.442mmol) and stirring 30min are added, DIPEA (0.771mL, 4.42mmol) and 2- is then added ((tetrahydrochysene -2H- pyrans -4- bases) epoxide) pyrimidine -4- amine (345mg, 1.766mmol), then stirs 16h at 80 DEG C.(TLC systems System:Rf-0.4,40%EtOAc- petroleum ethers).So that reactant mixture reaches room temperature and is quenched with water (30ml), ethyl acetate is used Extract (2x50ml).The organic layer of merging is through anhydrous sodium sulfate drying and is concentrated under reduced pressure, and obtains crude compound.Crude product is through fast Fast column chromatography purifies (silica gel：100-200 mesh, eluant, eluent：40% ethyl acetate/hexane) and by it again by preparation HPLC (the condition of purifying:Post：XBridge C 18 (75X4.6mm, 3.5 μ) mobile phase:A:0.01M ammonium hydrogen carbonate B:CAN gradients：When Between/%B:0/5,0.8/5,5/50,8/95,12/95,12.1/5,15/5 column temperature：Environment temperature, flow velocity:1.0ml/min dilution Agent：ACN), the chloro- N- of desired product (4S) -8- (2- ((tetrahydrochysene -2H- pyrans -4- bases) epoxide) pyrimidine-4-yl) -7- is obtained (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine (111.4mg, 0.198mmol, 22.46% yield), it is pale solid.LCMS(m/z):561.1[M+H]⁺, Rt= 2.70min。

¹H NMR(400MHz,CDCl₃):δ ppm 12.94 (s, 1H), 8.33 (d, J=5.70Hz, 1H), 8.07 (dd, J= 2.52,1.86Hz, 2H), 7.79-7.73 (m, 1H), 7.73-7.64 (m, 3H), 5.64 (dd, J=5.92,3.07Hz, 1H), 4.87 (tt, J=7.56,3.73Hz, 1H), 3.97-3.85 (m, 2H), 3.51 (ddt, J=11.46,7.62,3.51, 3.51Hz, 2H), 3.36-3.10 (m, 3H), 3.02 (dd, J=12.28,3.29Hz, 1H), 2.35 (dddd, J=14.14, 10.03,6.08,3.95Hz, 1H), 2.08 (dt, J=14.25,6.91Hz, 1H), 1.91-1.80 (m, 2H), 1.77-1.62 (m,2H)。

Embodiment 56

Synthesize (4S) -8- chloro- N- (6- ((R) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (6- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaIn the solution of stirring of -5 (the 2H)-formamides (9.1g, 15.42mmol) in methanol (100mL) add HCl (4.69mL, 154mmol).Reactant mixture is stirred into 1h.(TLC systems:Rf-0.4, net EtOAc) in room temperature and it is concentrated under reduced pressure, obtain remaining Thing.Residue is neutralized with saturated sodium bicarbonate solution, the solid product of acquisition is filtered and dried, desired product is obtained The chloro- N- of (4S) -8- (6- ((R) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- two Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamide (5.75g, 10.39mmol, 67.3% Yield), it is pale solid.LCMS(m/z):550.16[M+H]⁺, Rt=4.23min.

¹H NMR(400MHz,DMSO-d₆):δppm 12.44(s,1H),8.07-7.99(m,2H),7.90-7.80(m, 3H), 7.69-7.57 (m, 2H), 6.43 (dd, J=7.45,1.10Hz, 1H), 5.49 (dd, J=5.92,3.07Hz, 1H), 4.76 (d, J=4.82Hz, 1H), 4.53 (t, J=5.59Hz, 1H), 3.76-3.59 (m, 3H), 3.37-3.30 (m, 3H), 3.17-3.08 (m, 2H), 2.98 (dd, J=12.06,3.29Hz, 1H), 2.33-2.19 (m, 1H), 2.05-1.88 (m, 1H).

Embodiment 57

Synthesize (4S) -8- chloro- N- (4- ((S) -2,3- dihydroxy propoxyl group) pyrimidine -2-base) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (4- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyrimidine -2-base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaIn the solution of stirring of -5 (the 2H)-formamides (200mg, 0.338mmol) in methanol (8mL) add hydrochloric acid (1.0mL, 11.85mmol), 1h is then stirred at room temperature.(TLC systems:Net ethyl acetate, Rf:0.2).Reactant mixture is concentrated in vacuo And by residue saturation NaHCO₃Solution neutralizes and filters the solid of acquisition, then with pentane (2x10mL), ether (2x10mL) is washed, and obtains the chloro- N- of desired product (4S) -8- (4- ((S) -2,3- dihydroxy propoxyl group) pyrimidine -2-base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first Acid amides (92mg, 0.162mmol, 47.8% yield), it is pale solid.LCMS(m/z):551.13[M+H]⁺,R_t= 1.94min。

¹H NMR(400MHz,CDCl₃):δ ppm 13.40 (s, 1H), 8.19 (d, J=5.70Hz, 1H), 8.15 (s, 1H), 8.07 (d, J=7.45Hz, 1H), 7.74 (s, 1H), 7.68 (s, 1H), 7.66-7.59 (m, 1H), 6.42 (d, J=5.70Hz, 1H), 5.73 (dd, J=5.92,3.29Hz, 1H), 4.65 (dd, J=12.17,5.15Hz, 1H), 4.48 (dd, J=12.06, 4.60Hz, 1H), 4.25 (d, J=6.36Hz, 1H), 3.97-3.88 (m, 1H), 3.67-3.59 (m, 2H), 3.41-3.10 (m, 4H), 2.99 (dd, J=12.39,3.40Hz, 1H), 2.33 (qd, J=10.05,3.62Hz, 1H), 2.14-2.00 (m, 1H).

Embodiment 58

Synthesize (4S) -8- chloro- N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides, hydrochloride

At 0 DEG C to the chloro- N- of (4S) -8- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (3- (fluoroforms Base) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (100mg, 2M HCl diethyl ether solution (5mL, 0.182mmol) 0.182mmol) is added dropwise in the solution of the stirring in ether (10mL), goes through When 2min a period of time.Then reactant mixture is stirred into 2h at 30 DEG C.(TLC systems:5% ethanol/methylene, R_f: 0.1), then reactant mixture is evaporated and washed (2 × 20mL) with pentane, the chloro- N- of desired product (4S) -8- are obtained (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-carboxamide hydrochlorides (92mg, 0.155mmol, 85% yield), it is Pale solid.LCMS(m/z):550.16[M+H]⁺,R_t=2.20min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 12.76 (s, 1H), 8.11 (d, J=1.32Hz, 2H), 7.99-7.92 (m, 2H), 7.89-7.82 (m, 1H), 7.09 (d, J=1.32Hz, 1H), 6.66 (dd, J=6.03,1.64Hz, 1H), 5.46 (dd, J=5.59,2.96Hz, 1H), 4.23 (dd, J=10.52,4.38Hz, 1H), 4.11 (dd, J=10.52,6.14Hz, 1H), 3.82-3.73 (m, 1H), 3.43 (d, J=5.70Hz, 1H), 3.39-3.30 (m, 3H), 3.25-3.16 (m, 2H), 3.13-3.07 (m, 3H), 2.37-2.25 (m, 1H), 2.01 (dt, J=14.09,7.10Hz, 1H).

Embodiment 59

Synthesize (4S) -8- chloro- N- (6- (3- hydroxy-3-methyls butoxy) pyridine -2- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides:

In room temperature to the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diazaTEA is added in the solution of the stirring of (500mg, 1.472mmol) in THF (40mL) (1.026mL, 7.36mmol), triphosgene (262mg, 0.883mmol) simultaneously stirs 30min.Then add 4- ((6- aminopyridines- 2- yls) epoxide) -2- methyl butyl- 2- alcohol (578mg, 2.94mmol) and by reactant mixture 100 DEG C heat 16h.(TLC is eluted Agent system:100%EtOAc, Rf-0.5, UV activation).Reactant mixture is cooled to room temperature, then distribute in water (5mL) and Between EtOAc (15mL).Organic layer is separated, through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into crude product.Roughization Compound is through chromatogram purification (neutral alumina, eluant, eluent：30% ethyl acetate/hexane), obtain (4S) -8- chloro- N- (6- (3- hydroxyls Base -3- methylbutoxy groups) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (590mg, 1.049mmol, 71.3% yield), it is white solid LCMS(m/z):561.98[M]⁺,R_t=2.85min.

¹H NMR(400MHz,CDCl₃):δppm 12.51(s,1H),8.02(s,1H),7.83-7.98(m,1H),7.58- 7.76 (m, 4H), 7.58-7.76 (m, 1H), 6.36 (d, J=8.11Hz, 1H), 5.68 (dd, J=5.92,3.07Hz, 1H), 3.79-3.95 (m, 2H), 3.11-3.36 (m, 3H), 2.94-3.11 (m, 1H), 2.34 (br dd, J=9.98,4.06Hz, 1H), 2.21-2.29 (m, 1H), 1.95-2.21 (m, 1H), 1.72 (t, J=5.92Hz, 2H), 1.19 (s, 6H).

Embodiment 60

Synthesize (4S) -8- chloro- N- (2- ((R) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-carboxamide hydrochlorides:

At 0 DEG C to the chloro- N- of (4S) -8- (2- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridin-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaHCl/water solution is added in the solution of stirring of -5 (the 2H)-formamides (22.0g, 37.3mmol) in methanol (50mL) (22mL, 261mmol, 36%), is then stirred at room temperature 5h.After the completion of reaction, volatile matter is evaporated under reduced pressure, thick material is obtained. Solid is freezed with ethyl acetate (2x10mL), ether (10mL) grinding, is obtained the pure chloro- N- (2- of (4S) -8- by thick material ((R) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-carboxamide hydrochlorides (15.792g, 26.9mmol, 72.1% yield), it is Pale solid.LCMS(m/z):550.16[M+H]⁺,R_t=2.21min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 12.78 (s, 1H), 8.13-8.03 (m, 3H), 8.00 (d, J= 6.14Hz, 1H), 7.96-7.92 (m, 1H), 7.89-7.83 (m, 1H), 7.13 (brS, 3H), 6.71 (dd, J=6.14, 1.75Hz, 1H), 5.48 (dd, J=5.92,3.07Hz, 1H), 4.24 (dd, J=10.52,4.17Hz, 1H), 4.12 (dd, J= 10.52,6.36Hz,1H),3.83-3.76(m,1H),3.62-3.51(m,1H),3.51-3.41(m,5H),2.49-2.40(m, 1H),2.20-2.03(m,1H)。

Embodiment 61

Synthesize (4S) -8- chloro- N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridin-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaIn the solution of stirring of -5 (the 2H)-formamides (18.4g, 31.2mmol) in methanol (50mL) be added dropwise HCl (9.48mL, 312mmol), 5min time is lasted.Then reactant mixture is stirred into 2h at 30 DEG C.(TLC:Eluant, eluent：5%MeOH/DCM: R_f-0.5；UV activation) and evaporation solvent.Reactant mixture is neutralized and filtered with sodium bicarbonate solution the solid obtained, is used Ether (2x50mL) and pentane (2x50mL) washing, obtain the chloro- N- of desired product (4S) -8- (2- ((S) -2,3- dihydroxy Propoxyl group) pyridin-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (15.6g, 28.3mmol, 91% yield), it is pale solid.LCMS(m/z): 550.09[M+H]⁺,Rt:2.22min。

¹H NMR(400MHz,CDCl₃):δ ppm 12.77 (s, 1H), 7.98 (s, 1H), 7.93 (d, J=7.67Hz, 1H), 7.85 (d, J=5.92Hz, 1H), 7.80 (d, J=7.67Hz, 1H), 7.66-7.72 (m, 2H), 7.03 (d, J=1.32Hz, 1H), 6.70 (dd, J=5.81,1.64Hz, 1H), 5.65 (dd, J=5.48,2.85Hz, 1H), 4.43 (d, J=4.38Hz, 2H), 4.28 (d, J=4.60Hz, 1H), 3.97 (d, J=4.38Hz, 1H), 3.60-3.70 (m, 2H), 3.10-3.36 (m, 3H), 2.99-3.05 (m, 1H), 2.86 (s, 1H), 2.29-2.41 (m, 1H), 2.07 (dt, J=14.31,7.43Hz, 1H).

Embodiment 62

Synthesize (4S) -8- chloro- N- (6- ((R) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides, hydrochloride

At 0 DEG C to the chloro- N- of (4S) -8- (6- ((R) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (3- (fluoroforms Base) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (200mg, 2M HCl diethyl ether solution (3.63mL, 7.26mmol) 0.363mmol) is added in the solution of stirring in ether (5mL) and is stirred Mix 4h.(TLC eluant, eluents：100%EtOAc:R_f-0；UV activation).Reactant mixture is concentrated in vacuo and by residue with just Pentane grinds (2x10mL), obtains the chloro- N- of (4S) -8- (6- ((R) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (3- (three Methyl fluoride) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamide hydrochloric acid Salt (151mg, 0.249mmol, 68.7% yield), it is pale solid.LCMS(m/z):551.03[M+H]⁺,R_t= 2.18min。

¹H NMR(400MHz,DMSO-d₆):δppm 12.63(s,1H),8.86(s,1H),8.06-8.02(m,2H), 7.99(s,1H),7.93(s,1H),7.88-7.81(m,2H),5.66-5.30(m,1H),3.90-3.61(m,3H),3.48- 3.07(m,6H),2.45-2.23(m,1H),2.12-1.95(m,1H)

Embodiment 63

Synthesize (4S) -8- chloro- N- (4- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (4- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaHCl/water solution is added in the solution of stirring of -5 (the 2H)-formamides (8.0g, 13.56mmol) in methanol (80mL) (8.24mL, 271mmol, 36%), then stirs 2h at 0 DEG C.(TLC eluant, eluents：100%EtOAc:R_f-0.2；UV activation). Reactant mixture is alkalized (until pH-8-9) with saturated sodium bicarbonate solution and solvent is evaporated under reduced pressure.By residue distribution in water Between (40mL) and DCM (150mL).Organic layer is separated, through anhydrous sodium sulfate drying, filters and evaporates filter vacuum, will be thick Material obtains the chloro- N- of (4S) -8- (4- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- with triturated under ether (50mL) (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine (5.8g, 10.53mmol, 78% yield), it is white solid LCMS (m/z):550.09[M+H]⁺,R_t=1.96min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.93 (s, 1H), 8.20 (s, 1H), 8.12 (d, J=7.89Hz, 1H), 8.04 (d, J=5.70Hz, 1H), 7.77-7.71 (m, 2H), 7.68-7.58 (m, 2H), 6.53 (dd, J=5.81,2.30Hz, 1H), 5.64 (dd, J=5.81,3.18Hz, 1H), 4.19-4.08 (m, 3H), 3.88-3.79 (m, 1H), 3.79-3.66 (m, 1H), 3.37-3.11 (m, 3H), 3.01 (dd, J=12.28,3.29Hz, 1H), 2.58 (d, J=4.17Hz, 1H), 2.34 (dddd, J=14.17,10.00,5.97,4.06Hz, 1H), 2.15-1.98 (m, 2H).

Embodiment 64

Synthesize (4S) -8- chloro- N- (4- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-carboxamide hydrochlorides

At 0 DEG C to the chloro- N- of (4S) -8- (4- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (3- (fluoroforms Base) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (200mg, 0.364mmol) 2M HCl diethyl ether solution (3.64mL, 7.27mmol) is added in the solution of the stirring in ether (5mL) simultaneously Stir 4h.(TLC eluant, eluents：100%EtOAc:R_f-0；UV activation).Reactant mixture is concentrated in vacuo and residue is used Pentane grinds (2x10mL), obtains the chloro- N- of desired product (4S) -8- (4- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- Base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-carboxamide hydrochloride (178mg, 0.295mmol, 81% yield), it is pale solid.LCMS(m/z):550.13[M +H]⁺,R_t=1.95min.

¹H NMR(400MHz,DMSO-d₆):δppm 13.06(br s,1H),8.19-8.12(m,2H),8.08(br d,J =6.14Hz, 1H), 8.03 (s, 1H), 7.94-7.87 (m, 1H), 7.86-7.77 (m, 1H), 7.24 (br s, 1H), 6.86 (br D, J=3.73Hz, 1H), 5.49 (br dd, J=5.70,3.07Hz, 1H), 4.11 (dd, J=9.98,3.84Hz, 1H), 3.97 (dd, J=9.98,6.25Hz, 1H), 3.86-3.78 (m, 1H), 3.53-3.39 (m, 3H), 3.33-3.14 (m, 3H), 2.39- 2.28(m,1H),2.13-2.00(m,1H)。

Embodiment 65

Synthesize (4S) -8- chloro- N- (6- ((R) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (6- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaHCl/water solution is added in the solution of stirring of -5 (the 2H)-formamides (7.25g, 12.27mmol) in methanol (75mL) (7.45mL, 245mmol, 36%) and stirring 2h.(TLC eluant, eluents：100%EtOAc:R_f-0.2；UV activation).Reaction is mixed Compound is alkalized (until pH-8-9) with saturated sodium bicarbonate solution and is evaporated under reduced pressure solvent.By residue diluted with water (50mL) simultaneously It is extracted into dichloromethane (4x50mL).The organic extract of merging filters through anhydrous sodium sulfate drying and steams filter vacuum Hair, by thick material with triturated under ether (50mL), obtains the chloro- N- of (4S) -8- (6- ((R) -2,3- dihydroxy propoxyl group) pyrazine -2- Base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (5.2g, 9.38mmol, 76% yield), it is white solid.LCMS(m/z):551.10[M+H]⁺,R_t= 2.16min。

¹H NMR(400MHz,CDCl₃):δppm 12.76(s,1H),8.88(s,1H),8.00(s,1H),7.95-7.89 (m, 2H), 7.77-7.73 (m, 1H), 7.70-7.65 (m, 2H), 5.69 (dd, J=6.03,3.18Hz, 1H), 4.04-3.84 (m, 3H), 3.67-3.46 (m, 2H), 3.37-3.21 (m, 2H), 3.18-3.12 (m, 1H), 3.03 (dd, J=12.28, 3.29Hz, 1H), 2.58 (d, J=4.60Hz, 1H), 2.36 (dddd, J=14.20,10.03,6.03,3.95Hz, 1H), 2.14-2.03(m,2H)。

Embodiment 66

Synthesize (4S) -8- chloro- N- (2- ((R) -2,3- dihydroxy propoxyl group) pyrimidine -5- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to the chloro- N- of (4S) -8- (2- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyrimidine -5- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaHCl/water solution is added in the solution of stirring of -5 (the 2H)-formamides (250mg, 0.423mmol) in methanol (5mL) (0.357mL, 4.23mmol, 36%) and stirring 2h.(TLC eluant, eluents：100%EtOAc:R_f-0.2；UV activation).Will reaction Mixture is alkalized (until pH-8-9) with saturated sodium bicarbonate solution and is evaporated under reduced pressure solvent.By residue diluted with water (5mL) And (2x10mL) is extracted into dichloromethane.The organic extract of merging is filtered and by filter vacuum through anhydrous sodium sulfate drying Evaporation, by thick material with ether (10mL) and pentane (10mL) grinding, obtain the chloro- N- of desired product (4S) -8- (2- ((R) - 2,3- dihydroxy propoxyl group) pyrimidine -5- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (157mg, 0.278mmol, 65.8% yield), it is pale solid. LCMS(m/z):551.13[M+H]⁺,R_t=2.15min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.56 (s, 1H), 8.56 (s, 2H), 7.98 (s, 1H), 7.90 (d, J= 7.67Hz, 1H), 7.77 (br d, J=7.89Hz, 1H), 7.70-7.59 (m, 2H), 5.65 (dd, J=5.70,3.07Hz, 1H),4.51-4.38(m,2H),4.14-4.06(m,1H),3.81-3.65(m,2H),3.38-3.19(m,4H),3.07-3.00 (m, 1H), 2.41-2.26 (m, 2H), 2.09 (dt, J=14.31,7.21Hz, 1H)

Embodiment 67

Synthesize (4S) -8- chloro- N- (6- (3- hydroxy propyloxy groups) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

0 DEG C to the chloro- N- of (4S) -8- (6- (3- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) propoxyl group) pyridine -2- bases) - 7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first Added in the solution of stirring of the acid amides (250mg, 0.405mmol) in methanol (10mL) HCl/water solution (0.184mL, 6.07mmol), 4h is stirred.(TLC eluant, eluents：5%MeOH/DCM, Rf:0.3).Reactant mixture is concentrated in vacuo and by remnants Thing saturation NaHCO₃Basified (5mL).Gained solid is filtered, and (15mL) is ground with pentane, is dried under reduced pressure, obtains desired The chloro- N- of product (4S) -8- (6- (3- hydroxy propyloxy groups) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (61mg, 0.113mmol, 28.0% yield), It is pale solid.LCMS(m/z):534.14[M+H]⁺, Rt=2.62min.

¹H NMR(400MHz,CDCl₃):δppm 12.58(s,1H),7.89-8.03(m,2H),7.68-7.75(m,1H), 7.63-7.67 (m, 1H), 7.56-7.61 (m, 1H), 7.50-7.55 (m, 1H), 6.36 (dd, J=7.78,0.77Hz, 1H), 5.68 (dd, J=5.92,3.07Hz, 1H), 3.81-3.94 (m, 2H), 3.58 (q, J=5.70Hz, 2H), 3.19-3.36 (m, 2H), 3.11-3.18 (m, 1H), 3.01 (dd, J=12.28,3.29Hz, 1H), 2.27-2.40 (m, 1H), 2.02-2.17 (m, 2H), 1.72 (quin, J=5.81Hz, 2H).

Embodiment 68

Synthesize (4S) -8- chloro- N- (6- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-carboxamide hydrochlorides

At 0 DEG C to the chloro- N- of (4S) -8- (6- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (3- (fluoroforms Base) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (100mg, 2M HCl diethyl ether solution (8mL, 0.182mmol) 0.182mmol) is added in the solution of the stirring in methanol (5mL), then 2h is stirred at room temperature.(TLC eluant, eluents：Net ethyl acetate, Rf:0.1).Then it is concentrated in vacuo and grinds residue with pentane (10mLx3), obtains the chloro- N- of desired product (4S) -8- (6- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides Hydrochloride (40mg, 0.068mmol, 37.5% yield), it is pale solid.LCMS(m/z):550.09[M+H]⁺, Rt= 2.35min。

¹H NMR(400MHz,CDCl₃):δppm 12.16(s,1H),8.51(s,1H),8.02-7.95(m,2H),7.81 (d, J=7.67Hz, 1H), 7.74-7.69 (m, 1H), 7.62-7.56 (m, 1H), 7.50 (d, J=7.67Hz, 1H), 6.48 (d, J=7.67Hz, 1H), 5.91 (s, 1H), 4.02-3.47 (m, 11H), 2.66 (s, 1H), 2.47 (s, 1H).

Embodiment 69

Synthesize (4S) -8- chloro- N- (6- (2- hydroxyl-oxethyls) pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

0 DEG C to the chloro- N- of (4S) -8- (6- (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethyoxyl) pyrazine -2- bases) - 7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first HCl (1mL, 32.9mmol), Ran Hou are added in the solution of stirring of the acid amides (200mg, 0.331mmol) in methanol (10mL) 1h is stirred at room temperature.(TLC systems:Net ethyl acetate, Rf:0.3).Reactant mixture is concentrated in vacuo and by residue saturation NaHCO₃Solution is neutralized, and is filtered the solid obtained, is washed with water (20mLx3) and pentane (20mL × 2), obtain desired production The chloro- N- of thing (4S) -8- (6- (2- hydroxyl-oxethyls) pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (130mg, 0.247mmol, 74.7% yield), It is pale solid.LCMS(m/z):521.07[M+H]⁺, Rt=2.39min.

¹H NMR(400MHz,CDCl₃):δppm 12.75(s,1H),8.95(s,1H),7.98(s,1H),7.93-7.87 (m, 2H), 7.74-7.70 (m, 1H), 7.69 (s, 1H), 7.67-7.61 (m, 1H), 5.68 (dd, J=6.03,2.96Hz, 1H), 3.88-3.83 (m, 2H), 3.76-3.72 (m, 2H), 3.36-3.21 (m, 3H), 3.19-3.13 (m, 1H), 3.04 (dd, J= 12.28,3.07Hz,1H),2.41-2.31(m,1H),2.14-2.04(m,1H)。

Embodiment 70

Synthesize (4S) -8- chloro- N- (2- (2- hydroxyl-oxethyls) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

0 DEG C to the chloro- N- of (4S) -8- (2- (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethyoxyl) pyrimidine-4-yl) - 7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first HCl (0.5mL, 13.71mmol) is added in the solution of stirring of the acid amides (200mg, 0.331mmol) in methanol (10mL), so After 1h is stirred at room temperature.(TLC eluant, eluents：Net ethyl acetate, Rf:0.4).Reactant mixture is concentrated in vacuo and residue is used Saturation NaHCO₃Solution is neutralized, and filters the solid obtained, (20mLx3) is washed with pentane, the chloro- N- of (4S) -8- (2- (2- are obtained Hydroxyl-oxethyl) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (140mg, 0.267mmol, 81% yield), it is pale solid.LCMS(m/ z):521.04[M+H]⁺, Rt=2.33min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.04 (s, 1H), 8.34 (d, J=5.48Hz, 1H), 8.05-7.98 (m, 2H), 7.76-7.67 (m, 4H), 5.64 (dd, J=6.03,3.18Hz, 1H), 4.11-4.07 (m, 2H), 3.84-3.80 (m, 2H), 3.36-3.20 (m, 2H), 3.18-3.13 (m, 1H), 3.03 (dd, J=12.28,3.29Hz, 1H), 2.50 (t, J= 6.25Hz,1H),2.41-2.31(m,1H),2.13-2.04(m,1H)。

Embodiment 71

Synthesize (4S) -8- chloro- N- (6- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaIn the solution of stirring of -5 (the 2H)-formamides (8g, 13.56mmol) in methanol (70mL) add HCl (30mL, 987mmol), 2h is then stirred at room temperature.(TLC systems:Net ethyl acetate, Rf:0.3).Reactant mixture is concentrated in vacuo simultaneously By residue saturation NaHCO₃Solution is neutralized, and filters the solid obtained, (10mL × 2) are washed with pentane, obtain desired The chloro- N- of product (4S) -8- (6- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(6g, 10.87mmol, 80% are received -5 (2H)-formamides Rate), it is pale solid.LCMS(m/z):550.09[M+H]⁺, Rt=2.33min.

¹H NMR(400MHz,CDCl₃):δppm 12.61(s,1H),8.02-7.95(m,2H),7.76-7.72(m,1H), 7.69-7.64 (m, 2H), 7.58-7.53 (m, 1H), 7.51-7.48 (m, 1H), 6.42 (dd, J=7.89,0.66Hz, 1H), 5.68 (dd, J=5.92,3.07Hz, 1H), 3.97-3.88 (m, 2H), 3.75 (dq, J=10.14,5.10Hz, 1H), 3.54- 3.42 (m, 2H), 3.36-3.19 (m, 2H), 3.17-3.12 (m, 1H), 3.01 (dd, J=12.17,3.18Hz, 1H), 2.81 (d, J=5.92Hz, 1H), 2.38-2.28 (m, 2H), 2.13-2.04 (m, 1H).

Embodiment 72

Synthesize (4S) -8- chloro- N- (6- ((S) -2,3- dihydroxy propoxyl group) pyridin-3-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridin-3-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaAdd in the solution of stirring of -5 (the 2H)-formamides (250mg, 0.424mmol) in methanol (5mL) and tetrahydrofuran (5mL) Enter HCl (0.2mL, 6.58mmol), 3h is then stirred at room temperature.(TLC systems:Net ethyl acetate, Rf:0.1).By reaction mixing Thing is concentrated in vacuo and by residue saturation NaHCO₃Solution is neutralized and is extracted with ethyl acetate (30mL × 2).What is merged is organic Layer is washed with water (15mL × 2) and salt solution (20mL), through anhydrous sodium sulfate drying, is filtered and is concentrated under reduced pressure, and obtains thick material.Slightly Material grinds (15mL × 3) and filtering with pentane, obtains the chloro- N- of desired product (4S) -8- (6- ((S) -2,3- dihydroxy Propoxyl group) pyridin-3-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (175mg, 0.318mmol, 75% yield), it is pale solid.LCMS(m/z): 550.09[M+H]⁺, Rt=2.28min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.40 (s, 1H), 8.07 (d, J=2.41Hz, 1H), 7.99 (s, 1H), 7.93 (d, J=7.89Hz, 1H), 7.78-7.74 (m, 2H), 7.68-7.63 (m, 2H), 6.73 (d, J=8.99Hz, 1H), 5.66 (dd, J=5.92,3.07Hz, 1H), 4.42 (dd, J=4.60,1.10Hz, 2H), 4.02-3.93 (m, 2H), 3.72- 3.61 (m, 2H), 3.36-3.20 (m, 2H), 3.16-3.11 (m, 1H), 3.05-3.00 (m, 1H), 2.67 (t, J=6.47Hz, 1H), 2.38-2.29 (m, 1H), 2.09 (dt, J=14.25,7.13Hz, 1H).

Embodiment 73

Synthesize (4S) -8- chloro- N- (6- (3- hydroxy propyloxy groups) pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

It is sub- to the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- bridges in room temperature under a nitrogen Picoline simultaneously [2,3-b] [1,4] diazaThe solution of the stirring of (300mg, 0.883mmol) in tetrahydrofuran (15mL) Middle addition triphosgene (131mg, 0.442mmol) and stirring 30min.Into the reactant mixture add triethylamine (0.615mL, 4.42mmol) with 3- ((6- Aminopyrazine -2- bases) epoxide) propyl- 1- alcohol (194mg, 1.148mmol), then in 80 DEG C of stirrings 15h.(TLC systems:5% methanol/ethyl acetate.Rf values：0.3).(40mL) is diluted with water in reactant mixture and acetic acid second is used Ester extracts (2x50ml).The organic layer of merging through anhydrous sodium sulfate drying and is concentrated under reduced pressure with aqueous salt solu-tion (30mL), is obtained Obtain crude compound.Crude product is expected through flash column chromatography (100-200 silica gel, eluted with 1%MeOH/EtOAc) The chloro- N- of product (4S) -8- (6- (3- hydroxy propyloxy groups) pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(80mg, 0.144mmol, 16.33% are received -5 (2H)-formamides Rate), it is pale solid.LCMS(m/z):535.08[M+H]⁺, Rt=2.45min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.73 (s, 1H), 8.91 (s, 1H), 7.99 (s, 1H), 7.91 (d, J= 7.67Hz, 1H), 7.87 (s, 1H), 7.75-7.70 (m, 1H), 7.68 (s, 1H), 7.66-7.59 (m, 1H), 5.69 (dd, J= 5.92,3.07Hz, 1H), 3.97-3.86 (m, 2H), 3.67 (q, J=5.92Hz, 2H), 3.37-3.08 (m, 3H), 3.03 (dd, J=12.17,3.18Hz, 1H), 2.43-2.28 (m, 1H), 2.17-2.00 (m, 1H), 1.81 (q, J=5.97Hz, 2H), 1.67 (t, J=5.70Hz, 1H).

Embodiment 74

Synthesize (4S) -8- chloro- N- (2- (3- hydroxyl -2,2- dimethyl propylenes epoxide) pyrimidine-4-yl) -7- (3- (fluoroforms Base) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

It is sub- to the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- bridges in room temperature under a nitrogen Picoline simultaneously [2,3-b] [1,4] diazaAdded in the solution of (400mg, 1.177mmol) in tetrahydrofuran (20mL) Triphosgene (175mg, 0.589mmol) and stirring 30min.Into the reactant mixture add triethylamine (0.821mL, 5.89mmol) with 3- ((4- aminopyrimidine -2- bases) epoxide) -2,2- dimethyl propylene -1- alcohol (302mg, 1.531mmol), then 15h is stirred at 80 DEG C.(TLC systems:Ethyl acetate .Rf values：0.5).So that reactant mixture is cooled to room temperature and is diluted with water (20mL), is extracted with ethyl acetate (2x50ml).The organic layer of merging is dry through anhydrous sodium sulfate with aqueous salt solu-tion (20mL) It is dry and be concentrated under reduced pressure, obtain crude compound.Through flash column chromatography, (100-200 silica gel, uses 5%MeOH/ to crude product EtOAc elute), obtain the chloro- N- of desired product (4S) -8- (2- (3- hydroxyl -2,2- dimethyl propylenes epoxide) pyrimidine-4-yl) - 7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first Acid amides (120mg, 0.211mmol, 17.92% yield), it is pale solid.LCMS(m/z):563.12[M+H]⁺, Rt= 2.66min。

¹H NMR(400MHz,CDCl₃):δ ppm 13.06 (s, 1H), 8.32 (d, J=5.70Hz, 1H), 8.10 (s, 2H), 7.66-7.79 (m, 4H), 5.64 (dd, J=5.92,3.07Hz, 1H), 4.09-3.92 (m, 2H), 3.36-3.29 (m, 3H), 3.28-3.19 (m, 1H), 3.18-3.12 (m, 1H), 3.02 (dd, J=12.28,3.29Hz, 1H), 2.94 (brs, 1H), 2.41-2.30 (m, 1H), 2.07 (dt, J=14.36,7.07Hz, 1H), 0.97 (d, J=1.75Hz, 6H).

Embodiment 75

Synthesize (4S) -8- chloro- N- (6- ((S) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaHCl/water solution is added in the suspension of stirring of -5 (the 2H)-formamides (8.5g, 14.38mmol) in methanol (30mL) (50mL, 100mmol, 2M), is then stirred at room temperature 1h.(TLC systems:10% methanol/DCM.R_f:0.2).By reactant mixture Alkalized with saturated sodium bicarbonate solution.Gained white precipitate is filtered and dried, crude product is obtained.Purified on column chromatography is pure (silica gel uses 12%MeOH/CH for change₂Cl₂Elution), obtain the chloro- N- of (4S) -8- (6- ((S) -2,3- dihydroxy propoxyl group) pyrimidine - 4- yls) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (5.445g, 9.88mmol, 68.7% yield), it is white solid.LCMS (m/z)=551.1 [M+H]⁺, Rt=2.37min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.06 (s, 1H), 8.30 (d, J=0.88Hz, 1H), 8.04-8.21 (m, 2H), 7.85-7.94 (m, 2H), 7.69-7.85 (m, 1H), 7.39 (d, J=1.10Hz, 1H), 5.46 (dd, J=5.92, 3.07Hz, 1H), 4.95 (d, J=5.26Hz, 1H), 4.65 (t, J=5.70Hz, 1H), 4.34 (dd, J=10.85,4.06Hz, 1H), 4.19 (dd, J=10.96,6.58Hz, 1H), 3.74-3.86 (m, 1H), 3.35-3.54 (m, 2H), 3.22-3.28 (m, 1H),2.94-3.22(m,3H),2.15-2.32(m,1H),1.88-2.09(m,1H)。

Embodiment 76

Synthesize (4S) -8- chloro- N- (5- ((R) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamide

At 0 DEG C to the chloro- N- of (4S) -8- (5- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaIn the solution of stirring of -5 (the 2H)-formamides (200mg, 0.339mmol) in methanol (10mL) add hydrochloric acid (0.103mL, 3.39mmol), 2h is then stirred at room temperature.(TLC systems:10%MeOH/DCM, Rf:0.3).Reactant mixture is concentrated in vacuo And by residue NaHCO₃The aqueous solution is neutralized, and the solid of acquisition is filtered, and (2x10mL) is then washed with water, is expected The chloro- N- of product (4S) -8- (5- ((R) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (150mg, 0.271mmol, 80% yield), it is pale solid.LCMS(m/z):550.13[M+H]⁺, Rt=2.28min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.84 (s, 1H), 8.19 (s, 1H), 8.10 (d, J=7.89Hz, 1H), 8.03 (d, J=8.99Hz, 1H), 7.95 (d, J=3.07Hz, 1H), 7.73 (d, J=7.67Hz, 1H), 7.67-7.59 (m, 2H), 7.23 (d, J=3.07Hz, 1H), 5.66 (dd, J=6.14,3.07Hz, 1H), 4.15-4.08 (m, 1H), 4.08-4.02 (m, 2H), 3.89-3.82 (m, 1H), 3.79-3.74 (m, 1H), 3.34-3.12 (m, 3H), 3.01 (dd, J=12.28, 3.29Hz, 1H), 2.54 (d, J=4.17Hz, 1H), 2.38-2.28 (m, 1H), 2.15-2.04 (m, 1H), 1.94 (t, J= 5.70Hz,1H)。

Embodiment 77

Synthesize (4S) -8- chloro- N- (6- ((R) -2,3- dihydroxy propoxyl group) pyridin-3-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (6- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridin-3-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaIn the solution of stirring of -5 (the 2H)-formamides (350mg, 0.593mmol) in methanol (10mL) add HCl (0.180mL, 5.93mmol), 2h is then stirred at room temperature.(TLC systems:100% ethyl acetate, Rf values：0.2).Reactant mixture is used NaHCO₃Solution (10mL) is quenched and extracted (2x30mL) with DCM.The organic layer of merging is through anhydrous Na₂SO₄Dry, filtering simultaneously will Filtrate is evaporated, and obtains crude residue.Residue is ground into (3 × 10mL) with pentane, desired product (4S) -8- is obtained Chloro- N- (6- ((R) -2,3- dihydroxy propoxyl group) pyridin-3-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (160mg, 0.291mmol, 49.0% yield), its For white solid.LCMS(m/z):550.16[M+H]⁺, Rt=2.26min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.40 (s, 1H), 8.07 (d, J=2.85Hz, 1H), 7.99 (s, 1H), 7.93 (d, J=7.89Hz, 1H), 7.78-7.73 (m, 2H), 7.68-7.62 (m, 2H), 6.73 (d, J=8.77Hz, 1H), 5.66 (dd, J=5.92,3.07Hz, 1H), 4.42 (dd, J=4.60,2.41Hz, 2H), 4.02-3.91 (m, 2H), 3.72- 3.60 (m, 2H), 3.36-3.18 (m, 2H), 3.17-3.11 (m, 1H), 3.05-2.98 (m, 1H), 2.68 (t, J=6.14Hz, 1H), 2.34 (dddd, J=14.06,10.00,5.97,3.95Hz, 1H), 2.14-2.02 (m, 1H).

Embodiment 78

Synthesize (4S) -8- chloro- N- (5- ((S) -2,3- dihydroxy propoxyl group) pyridin-3-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (5- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridin-3-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaAdded in the solution of stirring of -5 (the 2H)-formamides (9g, 15.25mmol) in methanol (30mL) HCl/water solution (5mL, 165mmol), 5h is stirred.(TLC eluant, eluents：Mobile phase:10%MeOH/DCM；R_f:0.2；UV activation).Reactive material is true Sky concentration, obtains viscous brown oily thing, is dissolved in icy water (10mL), is neutralized with saturated sodium bicarbonate solution and uses 10% MeOH/DCM (2x50mL) is extracted.The organic layer of merging is dried over sodium sulfate with salt water washing (20mL), filters and depressurizes dense Contracting, obtains pale solid.Purified on column chromatography purifies to (silica gel uses 10%MeOH/CH₂Cl₂Elution), obtain (4S) -8- Chloro- N- (5- ((S) -2,3- dihydroxy propoxyl group) pyridin-3-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (6.5g, 11.81mmol, 77% yield), it is ash White solid.LC-MS(m/z):550.13[M+H]⁺, Rt=2.04min.

¹H NMR(400MHz,DMSO-d₆):δppm 12.55(s,1H),8.07-8.12(m,2H),7.90-7.96(m, 2H), 7.88 (s, 1H), 7.80-7.86 (m, 2H), 7.63 (t, J=2.30Hz, 1H), 5.47 (dd, J=5.81,2.96Hz, 1H), 4.97 (d, J=5.04Hz, 1H), 4.67 (t, J=5.70Hz, 1H), 4.00 (dd, J=9.43,3.73Hz, 1H), 3.76-3.88 (m, 2H), 3.41-3.48 (m, 2H), 3.22-3.32 (m, 1H), 3.10 (br d, J=11.84Hz, 2H), 2.95-3.03(m,1H),2.19-2.32(m,1H),1.91-2.01(m,1H)。

Embodiment 79

Synthesize (4S) -8- chloro- N- (6- (2- hydroxyl-oxethyls) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

0 DEG C to the chloro- N- of (4S) -8- (6- (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethyoxyl) pyridine -2- bases) - 7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first Added in solution of the acid amides (250mg, 0.414mmol) in methanol (10mL) HCl/water solution (0.6mL, 19.75mmol, 36%) and stirring 1h.(TLC eluant, eluents：100% ethyl acetate:R_f-0.2；UV activation).By reactant mixture saturated carbon The alkalization of sour hydrogen sodium solution is (until pH-8-9) and is evaporated under reduced pressure solvent.By residue diluted with water (10mL) and it is extracted into DCM (2x20mL).The organic extract of merging filters through anhydrous sodium sulfate drying and evaporates filtrate, by 10% acetic acid of thick material Ethyl ester/hexanes trituration, obtains the chloro- N- of (4S) -8- (6- (2- hydroxyl-oxethyls) pyridine -2- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (62mg, 0.117mmol, 28.4% yield), it is pale solid.LCMS(m/z):520.17[M+H]⁺,R_t=2.57min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.56 (s, 1H), 7.99 (s, 1H), 7.92 (d, J=7.45Hz, 1H), 7.75-7.69 (m, 1H), 7.69-7.58 (m, 3H), 7.57-7.50 (m, 1H), 6.41 (d, J=7.89Hz, 1H), 5.68 (dd, J=5.81,3.18Hz, 1H), 3.89-3.73 (m, 2H), 3.71-3.58 (m, 2H), 3.38-3.19 (m, 2H), 3.18-3.10 (m, 1H), 3.03 (dd, J=12.17,3.18Hz, 1H), 2.43-2.24 (m, 1H), 2.19-2.00 (m, 2H).

Embodiment 80

Synthesize (4S) -8- chloro- N- (2- ((S) -2,3- dihydroxy propoxyl group) pyrimidine -5- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Under a nitrogen at 0 DEG C to the chloro- N- of (4S) -8- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) Methoxyl group) pyrimidine -5- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diazaHCl/water solution is added in solution of -5 (the 2H)-formamides (300mg, 0.508mmol) in methanol (10mL) (1.2mL, 0.508mmol, 36%) and stirring 2h.(TLC eluant, eluents：100% ethyl acetate R_f-0.2；UV activation).At 0 DEG C Reactant mixture is alkalized (until pH-8-9) with saturated sodium bicarbonate solution and solvent is evaporated under reduced pressure.By residue diluted with water And stirring 15min.Then gained solid is dried by filtered on buchner funnel, obtains the chloro- N- of (4S) -8- (2- ((S) -2,3- bis- Hydroxy propyloxy group) pyrimidine -5- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (200mg, 0.358mmol, 70.5% yield), it is pale solid.LCMS(m/ z):551.17[M+H]⁺,R_t=3.86min.

¹H NMR(400MHz,DMSO-d₆):δppm 12.28(s,1H),8.60-8.46(m,2H),8.16-8.08(m, 2H), 7.95-7.86 (m, 2H), 7.84-7.74 (m, 1H), 5.45 (dd, J=5.70,3.07Hz, 1H), 4.91 (d, J= 5.26Hz, 1H), 4.62 (t, J=5.70Hz, 1H), 4.27 (dd, J=10.85,4.28Hz, 1H), 4.15 (dd, J=10.85, 6.47Hz, 1H), 3.79 (dq, J=10.74,5.55Hz, 1H), 3.43 (t, J=5.70Hz, 2H), 3.21-3.32 (m, 1H), 3.18-3.04 (m, 2H), 3.04-2.94 (m, 1H), 2.33-2.20 (m, 1H), 1.95 (dt, J=13.81,7.13Hz, 1H).

Embodiment 81

Synthesize (4S) -8- chloro- N- (5- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (5- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaAdded in solution of -5 (the 2H)-formamides (400mg, 0.678mmol) in methanol (15mL) HCl/water solution (0.016mL, 0.542mmol), 1h is then stirred at room temperature.(TLC systems:Rf:0.6,10%MeOH/DCM).By reactant mixture evaporation and Use NaHCO₃Solution is neutralized, and the solid of acquisition is filtered and washed (2x10mL) with ether, and obtaining half pure compound, (93% is pure Degree, is determined by LCMS).Half pure compound is purified into ((condition again by preparation HPLC：MP-A:10Mm ammonium hydrogen carbonate MP-B：Acetonitrile post：Xbridge (250x19) method-T/%B-0/10,1/10,10/55 flow velocitys：25ml/min eluant, eluents：ACN+ MeOH+THF), the chloro- N- of desired compound (4S) -8- (5- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- is obtained (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine (175mg, 0.318mmol, 46.9% yield), it is pale solid.LCMS(m/z):550.20 [M+H]+, Rt= 2.28min。

¹H NMR(400MHz,CDCl₃):δ ppm 12.84 (s, 1H), 8.19 (s, 1H), 8.10 (d, J=7.89Hz, 1H), 8.03 (d, J=8.99Hz, 1H), 7.95 (d, J=2.85Hz, 1H), 7.73 (d, J=7.89Hz, 1H), 7.65 (s, 1H), 7.65-7.60 (m, 1H), 7.25-7.22 (m, 1H), 5.66 (dd, J=5.92,3.07Hz, 1H), 4.14-4.02 (m, 3H), 3.88-3.82 (m, 1H), 3.79-3.72 (m, 1H), 3.36-3.12 (m, 3H), 3.01 (dd, J=12.17,3.18Hz, 1H), 2.58 (s, 1H), 2.38-2.28 (m, 1H), 2.09 (dt, J=14.14,6.96Hz, 1H), 1.98 (s, 1H).

Embodiment 82

Synthesize (4S) -8- chloro- N- (5- ((R) -2,3- dihydroxy propoxyl group) pyrimidine -2-base) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides:

At 0 DEG C to the chloro- N- of (4S) -8- (5- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyrimidine -2-base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaHCl/water solution is added in the solution of stirring of -5 (the 2H)-formamides (300mg, 0.508mmol) in methanol (15mL) (0.015mL, 0.508mmol, 36%), is then stirred at room temperature 1h. (TLC eluant, eluents：10%MeOH/EtOAc:R_f-0.1；UV Activation).Reactant mixture is alkalized (until pH-8-9) and concentrated with saturated sodium bicarbonate solution at 0 DEG C.Residue is used Water dilutes (8mL) and is extracted into DCM (2x25mL).The organic extract of merging is filtered and will filtered through anhydrous sodium sulfate drying Liquid is evaporated under reduced pressure, and obtains the chloro- N- of desired product (4S) -8- (5- ((R) -2,3- dihydroxy propoxyl group) pyrimidine -2-base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine (130mg, 0.229mmol, 45.1% yield), it is faint yellow solid.LCMS(m/z):551.10[M+H]⁺,R_t= 2.12min。

¹H NMR(400MHz,CDCl₃):δppm 13.21(s,1H),8.29(s,2H),8.15(s,1H),7.90-8.11 (m, 1H), 7.72 (br d, J=7.67Hz, 1H), 7.66 (s, 1H), 7.57-7.64 (m, 1H), 5.72 (dd, J=5.59, 3.18Hz,1H),4.00-4.22(m,3H),3.71-3.97(m,2H),3.08-3.37(m,3H),2.82-3.08(m,1H), 2.63(br s,1H),2.26-2.45(m,1H),1.96-2.19(m,2H)。

Embodiment 83

Synthesize (4S) -8- chloro- N- (5- ((S) -2,3- dihydroxy propoxyl group) pyrimidine -2-base) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Under a nitrogen at 0 DEG C to the chloro- N- of (4S) -8- (5- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) Methoxyl group) pyrimidine -2-base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diazaHCl/water solution is added in solution of -5 (the 2H)-formamides (300mg, 0.508mmol) in methanol (10mL) (1mL, 32.9mmol, 36%), is then stirred at room temperature 1h.(TLC eluant, eluents：5% methanol/DCM, Rf:0.3, UV activation). Saturation NaHCO is added into reactant mixture₃Solution (until pH-8-9), is then extracted into EtOAc (3x10mL).Merge Organic extract is through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain crude product.Thick material triturated under ether (2x10mL), obtains the chloro- N- of (4S) -8- (5- ((S) -2,3- dihydroxy propoxyl group) pyrimidine -2-base) -7- (3- (trifluoromethyl) Phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (250mg, 0.449mmol, 88% yield), it is pale solid.LCMS(m/z):551.13[M+H]⁺,R_t=2.10min.

¹H NMR(400MHz,CDCl₃):δppm 13.21(s,1H),8.32-8.27(m,2H),8.15(s,1H),8.06 (d, J=7.67Hz, 1H), 7.72 (d, J=7.89Hz, 1H), 7.67 (s, 1H), 7.63-7.56 (m, 1H), 5.72 (dd, J= 5.92,3.29Hz,1H),4.16-4.05(m,3H),3.90-3.82(m,1H),3.80-3.73(m,1H),3.36-3.11(m, 3H), 3.01 (dd, J=12.06,3.29Hz, 1H), 2.68 (br s, 1H), 2.38-2.27 (m, 1H), 2.10 (dt, J= 14.03,7.02Hz,2H)。

Embodiment 84

Synthesize (4S) -9- methyl -7- (2- picoline -4- bases)-N- (pyridin-3-yl) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (4S) -9- methyl -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diazaThree are added in the solution of the stirring of (400mg, 1.502mmol) in tetrahydrofuran (30ml) Phosgene (267mg, 0.901mmol) and DIPEA (0.787mL, 4.51mmol).Then reactant mixture is stirred at room temperature 30min.Pyridine -3- amine (212mg, 2.253mmol) is added after 30min, then 16h is stirred at 75 DEG C.(TLC systems：Only EtOAc,Rf:0.5).Then so that reactant mixture is cooled to room temperature, saturation NaHCO is poured into₃In solution (30mL), EtOAc is used Extract (2 × 100mL).The organic layer of merging is through anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain crude product.Thick material is passed through Flash column chromatography (neutral alumina, eluant, eluent：50%EtOAc/ petroleum ethers), obtain desired product (4S) -9- methyl - 7- (2- picoline -4- bases)-N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (294mg, 0.756mmol, 50.3% yield), it is white solid.LCMS(m/z):387.18[M +H]⁺,R_t=1.27min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.25 (s, 1H), 8.68-8.61 (m, 2H), 8.31 (dd, J=4.71, 1.42Hz, 1H), 8.19-8.11 (m, 1H), 7.58 (d, J=0.66Hz, 1H), 7.49 (dd, J=5.26,1.10Hz, 1H), 7.29-7.26 (m, 2H), 5.69 (dd, J=6.03,2.96Hz, 1H), 3.18 (t, J=7.45Hz, 2H), 3.13-2.99 (m, 2H),2.67(s,3H),2.51(s,3H),2.39-2.26(m,1H),2.12-2.00(m,1H)。

Embodiment 85

Synthesize (4S) -9- methyl-N- (pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (4S) -9- methyl -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diazaAdded in the solution of the stirring of (250mg, 0.783mmol) in tetrahydrofuran (30ml) Triphosgene (139mg, 0.470mmol) and triethylamine (0.109mL, 0.783mmol) and stirring 30min.After 30min, pyrrole is added Pyridine -2- amine (111mg, 1.174mmol), then stirs 16h at 75 DEG C.(TLC systems:EtOAc,Rf:0.6).Then so that anti- Answer mixture to be cooled to room temperature, pour into saturation NaHCO₃In solution (30mL), extracted with EtOAc (2 × 100mL).What is merged has Machine layer is through anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain crude product.Thick material is through flash column chromatography (neutral alumina Aluminium, eluant, eluent：20%EtOAc/ petroleum ethers), obtain desired product (4S) -9- methyl-N- (pyridine -2- bases) -7- (3- (three Methyl fluoride) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (70mg, 0.158mmol, 20.21% yield), it is white solid.LCMS(m/z):440.22[M+H]⁺,R_t=3.05min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.68 (br s, 1H), 8.46-8.30 (m, 3H), 8.17 (d, J= 8.33Hz, 1H), 7.74-7.59 (m, 3H), 7.36-7.24 (m, 1H), 7.02-6.92 (m, 1H), 5.71 (dd, J=5.59, 2.96Hz, 1H), 3.26-3.08 (m, 3H), 3.04-2.95 (m, 1H), 2.50 (s, 3H), 2.32 (td, J=13.76, 5.81Hz,1H),2.17-2.01(m,1H)。

Embodiment 86

Synthesize (4S) -9- methyl-N- (4- (2- Jia Ji oxazole -5- bases) pyridine -2- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (4S) -9- methyl -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diazaAdded in the solution of the stirring of (300mg, 0.939mmol) in tetrahydrofuran (30ml) Triphosgene (167mg, 0.564mmol) and DIPEA (0.820mL, 4.70mmol) and stirring 30min.Then 4- (2- methyl is added Oxazole -5- bases) pyridine -2- amine (247mg, 1.409mmol), then stirs 16h at 75 DEG C.(TLC systems:EtOAc,Rf: 0.3).Then reactant mixture is cooled to room temperature, pour into saturation NaHCO₃Solution (30mL), extracted with EtOAc (2 × 100mL).The organic layer of merging is through anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain crude product.Thick material is through quick post color Spectrum purifying (neutral alumina, eluant, eluent：40%EtOAc/ petroleum ethers), obtain desired product (4S) -9- methyl-N- (4- (2- Jia Ji oxazole -5- bases) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza- 5 (2H)-formamides (126mg, 0.241mmol, 25.7% yield), it is white solid.LCMS (m/z):521.29[M+H]⁺；R_t=3.13min.

¹H NMR(400MHz,CDCl₃):δppm 13.81(s,1H),8.51-8.29(m,4H),7.72-7.60(m,2H), 7.45 (s, 1H), 7.32 (s, 1H), 7.17 (dd, J=5.15,1.43Hz, 1H), 5.72 (dd, J=5.92,3.07Hz, 1H), 3.23-3.09 (m, 3H), 3.08-2.96 (m, 1H), 2.53 (s, 3H), 2.51 (s, 3H), 2.34 (td, J=13.76, 6.47Hz,1H),2.16-2.02(m,1H)。

Embodiment 87

Synthesize (4S)-N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -9- methyl -7- (3- (trifluoromethyl) Phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature to (4S)-N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine - 4- yls) -9- methyl -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneousIn the solution of stirring of -5 (the 2H)-formamides (400mg, 0.702mmol) in methanol (30mL) add hydrochloric acid (2mL, 23.70mmol) and stirring 1h.(TLC systems：Net EtOAc, Rf:0.3).After 1h, reactant mixture is concentrated under reduced pressure and saturation is used NaHCO₃(30mL) is quenched in solution, is extracted with 5%MeOH/DCM (2x50mL).The organic layer of merging through anhydrous sodium sulfate drying simultaneously It is concentrated under reduced pressure, obtains semi-solid, it is washed to (2x20mL) with pentane and dried, desired product (4S)-N- (2- are obtained ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -9- methyl -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (288mg, 0.538mmol, 77% yield), it is Pale solid.LCMS(m/z):530.29[M+H]⁺,R_t=2.32min.

¹H NMR(400MHz,CDCl₃):δppm 13.43(s,1H),8.05(s,1H),8.00-7.88(m,2H),7.78- 7.62 (m, 2H), 7.23 (s, 1H), 7.16 (s, 1H), 6.94 (d, J=4.38Hz, 1H), 5.67 (dd, J=5.26,2.85Hz, 1H), 4.45 (d, J=4.38Hz, 3H), 4.03-3.93 (m, 1H), 3.74-3.59 (m, 2H), 3.23-2.99 (m, 5H), 2.50 (s, 3H), 2.33 (td, J=13.54,6.25Hz, 1H), 2.13-1.99 (m, 1H).

Embodiment 88

Synthesize (4S) -8- chloro- N- (6- methyl isophthalic acid H- pyrazolos [3,4-b] pyridin-3-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In a nitrogen atmosphere in room temperature to the chloro- 7- of (4S) -8- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysenes -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diazaThe stirring of (600mg, 1.766mmol) in tetrahydrofuran (10mL) Triphosgene (524mg, 1.766mmol) and TEA (0.738mL, 5.30mmol) are added in solution, 30min is then stirred for.With 6- methyl isophthalic acid H- pyrazolos [3,4-b] pyridine -3- amine (523mg, 3.53mmol) is added thereto in room temperature afterwards, then at 75 DEG C Stir 15h.(TLC 5%MeOH DCM Rf:0.3；UV activation).(10mL) is diluted with water in reactant mixture and acetic acid is used Ethyl ester extracts (2 × 20ml).The organic layer of merging is washed with saturated brine solution, then through anhydrous sodium sulfate drying, is depressurized dense Contracting, obtains crude compound, it is purified into (silica gel by column chromatography:100-200 mesh, eluant, eluent：80%EtOAc/ petroleum ethers), Obtain the chloro- N- of desired product (4S) -8- (6- methyl isophthalic acid H- pyrazolos [3,4-b] pyridin-3-yl) -7- (3- (trifluoromethyl) Phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (243mg, 0.451mmol, 25.5% yield), it is white solid.LCMS(m/z):514.23[M+H]⁺, Rt=2.54min.

¹H NMR(400MHz,DMSO-d₆+ 1 drop TFA):δppm 8.54-8.36(m,1H),8.17-8.01(m,3H), 7.92 (d, J=7.7Hz, 1H), 7.80 (t, J=7.8Hz, 1H), 7.18-7.00 (m, 1H), 5.63 (dd, J=3.0,5.6Hz, 1H), 3.57-3.38 (m, 4H), 2.69-2.56 (m, 3H), 2.49-2.32 (m, 1H), 2.23 (td, J=7.1,13.9Hz, 1H)。

Embodiment 89

Synthesis (4S) -7- (3- (difluoromethyl) phenyl)-N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) - 8- methyl -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -7- (3- (difluoromethyl) phenyl)-N- (2- (((R) -2,2- dimethyl -1,3- dioxanes penta Alkane -4- bases) methoxyl group) pyridin-4-yl) -8- methyl -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneousIn the solution of stirring of -5 (the 2H)-formamides (0.2g, 0.363mmol) in methanol (10mL) be added dropwise HCl (5.51 μ L, 0.181mmol), 5min time is lasted.Then reactant mixture is stirred into 2h (TLC eluant, eluents at 30 DEG C：5%MeOH/DCM: R_f-0.3；UV activation).After 2h, reactant mixture is concentrated in vacuo and by residue NaHCO₃The aqueous solution is neutralized and will obtained Solid filter and use ether (2x50mL), pentane (2x50mL) washing, obtain desired product (4S) -7- (3- (difluoro first Base) phenyl)-N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -8- methyl -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (110mg, 0.215mmol, 59.2% yield), it is canescence Solid.LCMS(m/z):512.25[M+H]⁺, Rt=1.95min.

¹H NMR(400MHz,DMSO-d₆):δppm 13.28(s,1H),7.88-7.80(m,3H),7.74-7.70(m, 2H), 7.61 (s, 1H), 7.29-7.00 (m, 2H), 6.52 (dd, J=5.70,1.75Hz, 1H), 5.44 (dd, J=5.92, 3.29Hz, 1H), 4.85 (s, 1H), 4.58 (s, 1H), 4.20 (dd, J=10.74,4.60Hz, 1H), 4.08 (dd, J= 10.85,6.25Hz, 1H), 3.76 (d, J=5.04Hz, 1H), 3.42 (s, 2H), 3.26-3.17 (m, 1H), 3.14-3.05 (m, 2H), 2.94 (dd, J=12.06,3.29Hz, 1H), 2.31 (s, 3H), 2.26-2.20 (m, 1H), 1.95-1.85 (m, 1H).

Embodiment 90

Synthesize (4S) -8- chloro- N- (2- ((R) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (1- ethyl -1H- pyrroles Azoles -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamide

Under a nitrogen in room temperature to the chloro- N- of (4S) -8- (2- (((S) -2,2- dimethyl -1,3- dioxolane -4- Base) methoxyl group) pyridin-4-yl) -7- (1- ethyl -1H- pyrazoles -4- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza- 5 (2H)-formamides (400mg, 0.741mmol) add HCl in the solution in methanol (5mL) (2mL, 65.8mmol) and stir 2h.(5% methanol of TLC systems/DCM.Rf values：0.1).Reactant mixture is concentrated, residue Saturation NaHCO is used at 0 DEG C₃It is basified.Gained solid is filtered and dried, crude compound is obtained, solid is ground with ether Grind (10mL), obtain the chloro- N- of (4S) -8- (2- ((R) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (1- ethyl -1H- pyrroles Azoles -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (230mg, 0.458mmol, 61.8% yield), it is pale solid.LCMS(m/z):500.26[M+H]⁺, Rt=1.75min.

¹H NMR(400MHz,DMSO-d6):δ ppm 12.66 (s, 1H), 8.43 (s, 1H), 8.01 (t, J=2.7Hz, 2H), 7.74 (s, 1H), 7.10-6.87 (m, 2H), 5.41 (br d, J=3.1Hz, 1H), 4.86 (d, J=5.0Hz, 1H), 4.59 (br t, J=5.7Hz, 1H), 4.40-4.19 (m, 3H), 4.19-3.96 (m, 1H), 3.93-3.65 (m, 1H), 3.43 (br t, J=5.7Hz, 2H), 3.22 (br s, 1H), 3.16-3.00 (m, 2H), 2.93 (brdd, J=3.1,12.1Hz, 1H), 2.37-2.08 (m, 1H), 1.93 (brdd, J=6.4,13.8Hz, 1H), 1.46 (t, J=7.2Hz, 3H).

Embodiment 91

Synthesize (4S) -8,9- dimethyl -7- (2- picoline -4- bases)-N- (pyridin-3-yl) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

In a nitrogen atmosphere room temperature to (4S) -8,9- dimethyl -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysenes - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaThe stirring of (300mg, 1.070mmol) in THF (30mL) Pyridin-3-yl phenyl carbamate (688mg, 3.21mmol) is added in solution, DMAP (392mg, 3.21mmol) is subsequently added into And stir 16h at 65 DEG C.(TLC eluant, eluents：100%EtOAc:R_f-0.2；UV activation).Reactant mixture is cooled to room Temperature, concentration, by residue distribution between water (20mL) and EtOAc (50mL).Organic layer is separated, through anhydrous Na₂SO₄Dry, mistake Filter and evaporate filtrate, obtain crude compound.Purified on column chromatography is purified into (neutral alumina, eluant, eluent：70% EtOAc/ hexanes), obtain desired product (4S) -8,9- dimethyl -7- (2- picoline -4- bases)-N- (pyridin-3-yl) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (238mg, 0.590mmol, 55.1% yield), it is white solid.LCMS(m/z):401.12[M+H]⁺,R_t=1.35min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.19 (s, 1H), 8.65 (d, J=5.04Hz, 1H), 8.42 (br s, 1H), 8.24 (br d, J=4.17Hz, 1H), 8.03-7.94 (m, 1H), 7.29 (s, 1H), 7.23-7.16 (m, 2H), 5.64 (dd, J=5.81,2.96Hz, 1H), 3.25-3.14 (m, 2H), 3.12-3.00 (m, 2H), 2.67 (s, 3H), 2.47 (s, 3H), 2.38-2.28(m,1H),2.24(s,3H),2.12–1.99(m,1H)。

Embodiment 92

Synthesize (4S) -8- cyano group -7- (2- picoline -4- bases)-N- (pyridine -2- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the bromo- 7- of (4S) -8- (2- picoline -4- bases)-N- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (300mg, 0.665mmol), Zn (CN)₂(234mg, 1.994mmol) with Zn (OAc)₂The solution of the degassing of (73.2mg, 0.399mmol) in N,N-dimethylformamide (12mL) Middle addition solid Pd₂(dba)₃(122mg, 0.133mmol) and DPPF (147mg, 0.266mmol).Then reactant mixture is existed 100 DEG C of stirring 15h.(TLC systems；Rf-0.4,5% ethanol/methylenes).So that reactant mixture is cooled to room temperature and uses water Dilute (30mL), be extracted with ethyl acetate (3 × 50mL).The organic layer of merging is through anhydrous Na₂SO₄Dry, filter and depressurize dense Contracting, obtains crude compound.It is passed through through flash column chromatography (100-200 mesh) and further preparative by crude compound HPLC purifies (condition：MP-A:5mM ammonium hydrogen carbonate (aqueous solution) MP-B：Acetonitrile post：The μ sides of KROMOSIL C18 (250x21.2) 10 Method：0/50,12/50,12.5/100,18/100,18.1/50 flow velocity：19ml/min eluant, eluents：MeOH+THF+ACN), it must expire Product (4S) -8- cyano group -7- (2- picoline -4- bases)-N- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles of prestige Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (85mg, 0.211mmol, 31.8% yield), it is solid for canescence Body.LCMS(m/z):398.06[M+H]⁺,R_t=1.97min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.92 (s, 1H), 8.71 (d, J=5.26Hz, 1H), 8.34-8.29 (m, 1H), 8.17-8.12 (m, 1H), 7.96 (d, J=1.53Hz, 1H), 7.86-7.79 (m, 2H), 7.74-7.67 (m, 1H), 7.03 (ddd, J=7.29,4.88,0.99Hz, 1H), 5.71 (dd, J=5.81,2.96Hz, 1H), 3.33-3.22 (m, 2H), 3.16-3.02(m,2H),2.75(s,3H),2.44-2.31(m,1H),2.18-2.05(m,1H)。

Embodiment 93

Synthesize (4S) -8- chloro- N- (6- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to the chloro- N- of (4S) -8- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaAdded in the solution of stirring of -5 (the 2H)-formamides (250mg, 0.466mmol) in methanol (10mL) 3M HCl (10mL, 30.0mmol).1h is stirred at room temperature in gained mixture.(TLC5%MeOH/DCM Rf:0.2；UV activation) and by reactant Matter saturated sodium bicarbonate solution (10mL) alkalizes, and is extracted with DCM (2X20mL).The organic layer aqueous salt solu-tion of merging (10mL), through anhydrous sodium sulfate drying, filters and is concentrated under reduced pressure, and obtains brown solid, and it is ground with pentane (10mL) and Vacuum drying, obtains the chloro- N- of desired product (4S) -8- (6- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (2- Picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (115mg, 0.220mmol, 47.3% yield), it is faint yellow solid.LCMS(m/z):497.1[M+H]⁺, Rt= 1.54min。

¹H NMR(400MHz,DMSO-d₆):δ ppm 12.44 (s, 1H), 8.65 (d, J=5.04Hz, 1H), 7.85 (s, 1H), 7.69-7.57 (m, 2H), 7.54-7.45 (m, 2H), 6.44 (dd, J=7.67,0.88Hz, 1H), 5.48 (dd, J= 5.70,2.85Hz, 1H), 4.74 (d, J=4.60Hz, 1H), 4.67-4.44 (m, 1H), 3.81-3.58 (m, 3H), 3.46- 3.38(m,2H),3.32-3.25(m,1H),3.19-3.03(m,2H),3.04–2.83(m,1H),2.49(s,3H),2.36- 2.05 (m, 1H), 1.95 (dt, J=13.76,7.04Hz, 1H).

Embodiment 94

Synthesize (4S) -8- chloro- N- (2- ((S) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides:

At 0 DEG C to the chloro- N- of (4S) -8- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyrimidine-4-yl) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaHCl/water solution is added in the solution of stirring of -5 (the 2H)-formamides (150mg, 0.279mmol) in methanol (5mL) (0.5mL, 16.46mmol), is then stirred at room temperature 1h.(TLC eluant, eluents：10%MeOH/DCM, R_f=0.3；UV activation). Reactant mixture is concentrated under reduced pressure, and residue is alkalized (until pH-8-9) with saturated sodium bicarbonate solution and water layer is used DCM extracts (2 × 40mL).The organic layer of merging is through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain crude product.It is rough Compound with triturated under ether (2x15mL), obtain the chloro- N- of (4S) -8- (2- ((S) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) - 7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first Acid amides (125mg, 0.241mmol, 87% yield), it is pale solid.LCMS(m/z):498.10[M+H]⁺, Rt= 1.43min。

¹H NMR(400MHz,CDCl₃):δ ppm 13.01 (s, 1H), 8.68 (d, J=5.04Hz, 1H), 8.34 (d, J= 5.70Hz, 1H), 7.74 (d, J=5.48Hz, 1H), 7.69 (s, 1H), 7.57 (s, 1H), 7.52 (br d, J=5.26Hz, 1H), 5.63 (br dd, J=5.59,2.74Hz, 1H), 3.93-4.09 (m, 3H), 3.54-3.75 (m, 4H), 3.19-3.37 (m, 2H), 3.10-3.18 (m, 1H), 2.99-3.08 (m, 1H), 2.69 (s, 3H), 2.28-2.43 (m, 1H), 2.07 (dt, J= 14.41,7.15Hz,1H)。

Embodiment 95

Synthesize (4S) -8- chloro- N- (6- ((R) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (6- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyrimidine-4-yl) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaHCl is added in the solution of stirring of -5 (the 2H)-formamides (0.12g, 0.223mmol) in DCM (5mL) and methanol (5mL) (in 0.558mL, 2.230mmol, 4M dioxanes) and stirring 4h.(TLC eluant, eluents：10%MeOH/EtOAc:R_f-0.3；UV Activation).By reactant mixture by adding saturated sodium bicarbonate solution alkalization (until pH=8-9), then concentrate.By remnants Thing is diluted with water (10mL) and is extracted into EtOAc in (2x25mL).The organic extract of merging is through anhydrous Na₂SO₄Dry, mistake Filter and evaporate filtrate, obtain crude product.Crude compound purifies through column chromatography and (neutral alumina is used, with 50% acetic acid second Ester/Hex), obtain the chloro- N- of (4S) -8- (6- ((R) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (2- methyl pyrroles Pyridine -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (0.09g, 0.174mmol, 78% yield), it is pale solid.LCMS(m/z):498.07[M+H]⁺,R_t=1.40min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.02 (s, 1H), 8.62 (d, J=5.26Hz, 1H), 8.46 (s, 1H), 7.88 (s, 1H), 7.61-7.73 (m, 2H), 7.41 (s, 1H), 5.45 (br d, J=2.63Hz, 1H), 4.74-5.14 (m, 2H), 4.35 (br dd, J=10.85,4.06Hz, 1H), 4.21 (br dd, J=10.74,6.36Hz, 1H), 3.76- 3.86 (m, 1H), 3.43 (br d, J=5.04Hz, 2H), 3.19-3.26 (m, 1H), 3.06-3.15 (m, 2H), 2.97 (br Dd, J=12.06,2.85Hz, 1H), 2.61 (s, 3H), 2.20-2.28 (m, 1H), 1.91-2.05 (m, 1H).

Embodiment 96

Synthesize (4S) -8- chloro- N- (2- ((R) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

0 DEG C in a nitrogen atmosphere to the chloro- N- of (4S) -8- (2- (((S) -2,2- dimethyl -1,3- dioxolane - 4- yls) methoxyl group) pyrimidine-4-yl) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diazaAdded in the solution of stirring of -5 (the 2H)-formamides (250mg, 0.465mmol) in methanol (10mL) HCl/water solution (1.0ml, 4.00mmol).Reactant mixture is stirred into 1h at 25 DEG C.(TLC eluant, eluents：5% methanol/DCM, Rf: 0.3, UV activation).Solvent is evaporated under reduced pressure, is diluted with water, uses saturation NaHCO₃It is basified (until pH:8-9) and use dichloro Methane extracts (3 × 10mL).The organic layer of merging is with aqueous salt solu-tion (10mL), through anhydrous Na₂SO₄Dry, filter and steam Hair, obtains crude compound.Thick material with triturated under ether (10mL), obtain the chloro- N- of desired product (4S) -8- (2- ((R) -2, 3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza- 5 (2H)-formamides (125mg, 0.247mmol, 53.3% yield), it is pale solid. LCMS(m/z):498.07[M+H]⁺,R_t=1.44min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.02 (s, 1H), 8.68 (d, J=5.04Hz, 1H), 8.34 (d, J= 5.70Hz, 1H), 7.75 (d, J=5.70Hz, 1H), 7.69 (s, 1H), 7.56 (s, 1H), 7.51 (d, J=4.17Hz, 1H), 5.63 (dd, J=5.81,3.18Hz, 1H), 4.07-3.97 (m, 3H), 3.70 (d, J=4.60Hz, 2H), 3.60-3.10 (m, 5H),3.08–2.99(m,1H),2.68(s,3H),2.41-2.29(m,1H),2.07(m,1H)。

Embodiment 97

Synthesize (4S) -8- chloro- N- (6- ((R) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (6- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaHCl/water solution is added in the solution of stirring of -5 (the 2H)-formamides (180mg, 0.335mmol) in methanol (10mL) (0.2mL, 6.58mmol) and reactant mixture is stirred into 2h at 0 DEG C.(TLC eluant, eluents：5%MeOH/DCM；Rf values：0.3；UV Activation).Reactant mixture is concentrated in vacuo, at 0 DEG C by the residue of acquisition saturation NaHCO₃Solution neutralizes (20mL) simultaneously Extracted with DCM (2 × 40mL).The organic layer of merging is through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain rough chemical combination Thing.The compound purifies (30mL) through pentane, obtains the chloro- N- of desired product (4S) -8- (6- ((R) -2,3- dihydroxy third Epoxide) pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine- 5 (2H)-formamides (116mg, 0.228mmol, 68.3% yield), it is pale solid.LCMS(m/z): 498.07[M+H]⁺, Rt=1.44min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.70 (s, 1H), 9.00 (s, 1H), 8.65 (d, J=5.26Hz, 1H), 7.93 (s, 1H), 7.68 (s, 1H), 7.59-7.53 (m, 1H), 7.49-7.42 (m, 1H), 5.67 (dd, J=5.92,3.07Hz, 1H),4.01-3.91(m,1H),3.80-3.50(m,4H),3.39-3.20(m,2H),3.20-3.09(m,1H),3.09-2.99 (m, 1H), 2.71-2.58 (m, 3H), 2.47-2.26 (m, 1H), 2.08 (dt, J=14.09,7.10Hz, 1H).

Embodiment 98

Synthesize (4S) -8- chloro- N- (6- ((S) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to the chloro- N- of (4S) -8- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza4M HCl are added in the solution of stirring of -5 (the 2H)-formamides (200mg, 0.372mmol) in 1,4- dioxanes (5.0mL) Dioxane solution (0.929mL, 3.72mmol), in mutually synthermal stirring 4h (TLC:Eluant, eluent：Net ethyl acetate, Rf: 0.3).By reactant mixture distribution in saturation NaHCO₃Between the aqueous solution (10mL) and EtOAc (30mL).Organic layer is separated, so By anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into crude product.Crude compound is through flash column chromatography (silica gel: 100-200 mesh, eluant, eluent：60% ethyl acetate/hexane), obtain the chloro- N- of desired product (4S) -8- (6- ((S) -2,3- bis- Hydroxy propyloxy group) pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (120mg, 0.240mmol, 64.7% yield), it is white solid.LCMS(m/ z):498.07[M+H]⁺, Rt=1.44min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 12.70 (s, 1H), 8.86 (s, 1H), 8.65 (d, J=5.04Hz, 1H), 7.94 (s, 1H), 7.89 (s, 1H), 7.55 (s, 1H), 7.49 (s, 1H), 5.49 (dd, J=5.70,2.85Hz, 1H), 4.88 (d, J=5.04Hz, 1H), 4.65 (t, J=5.70Hz, 1H), 3.88-3.77 (m, 2H), 3.75-3.67 (m, 1H), 3.39 (t, J=5.81Hz, 2H), 3.32-3.30 (m, 1H), 3.16-3.09 (m, 2H), 2.99 (dd, J=11.95,3.18Hz, 1H),2.55(s,3H),2.30-2.20(m,1H),2.04-1.81(m,1H)。

Embodiment 99

Synthesize (4S) -8- chloro- N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (6- methoxyl groups -5- (three Methyl fluoride) pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine

At 0 DEG C to the chloro- N- of (4S) -8- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridin-4-yl) -7- (6- methoxyl groups -5- (trifluoromethyl) pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diazaAdd in the solution of stirring of -5 (the 2H)-formamides (330mg, 0.531mmol) in methanol (5mL) Enter HCl/water solution (0.448mL, 5.31mmol) and stir 2h.(TLC eluant, eluents：100%EtOAc:R_f-0.2；UV activation). Reactant mixture is alkalized (until pH-8-9) with saturated sodium bicarbonate solution and solvent is evaporated under reduced pressure.By residue diluted with water (5mL) and it is extracted into dichloromethane in (2x10mL).The organic extract of merging is filtered and will filtered through anhydrous sodium sulfate drying Liquid is evaporated in vacuo, and obtains the chloro- N- of desired product (4S) -8- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (6- methoxyl groups -5- (trifluoromethyl) pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (264mg, 0.453mmol, 85% yield), it is pale solid.LCMS(m/z):581.19[M +H]⁺,R_t=2.24min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.85-12.54 (m, 1H), 8.77 (d, J=2.2Hz, 1H), 8.24 (d, J=2.0Hz, 1H), 7.90 (d, J=5.9Hz, 1H), 7.68 (s, 1H), 7.02 (d, J=1.5Hz, 1H), 6.78 (dd, J =1.9,5.8Hz, 1H), 5.65 (dd, J=3.1,5.9Hz, 1H), 4.48-4.40 (m, 2H), 4.24 (br s, 1H), 4.16 (s, 3H), 3.98 (br t, J=4.5Hz, 1H), 3.71-3.57 (m, 2H), 3.35-3.18 (m, 2H), 3.16-3.08 (m, 1H), 3.06-2.95 (m, 1H), 2.85 (br t, J=6.4Hz, 1H), 2.44-2.25 (m, 1H), 2.07 (td, J=7.1, 14.3Hz,1H)。

Embodiment 100

Synthesize (4S) -8- chloro- 7- (1- cyclopropyl -1H- pyrazoles -4- bases)-N- (2- ((R) -2,3- dihydroxy propoxyl group) pyrroles Pyridine -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- 7- of (4S) -8- (1- cyclopropyl -1H- pyrazoles -4- bases)-N- (2- (((S) -2,2- dimethyl -1,3- Dioxolane -4- bases) methoxyl group) pyridin-4-yl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two AzepineHCl is added in the solution of stirring of -5 (the 2H)-formamides (250mg, 0.453mmol) in methanol (5mL) (0.069mL, 2.264mmol), is then stirred at room temperature 1h.(TLC systems:Rf-0.1,EtOAc).Reactant mixture is concentrated, Residue is neutralized with saturated sodium bicarbonate solution.Gained solid is filtered and dried, (4S) -8- chloro- 7- (1- rings third are obtained Base -1H- pyrazoles -4- bases)-N- (2- ((R) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (175mg, 0.337mmol, 74.4% yield), it is greyish white Color solid.LCMS(m/z):512.18[M+H]⁺.Rt=1.79min.

¹H NMR(400MHz,CDCl₃):δppm 12.80(s,1H),8.10(s,1H),8.02(s,1H),7.97(s, 1H), 7.59 (s, 1H), 7.13-7.01 (m, 2H), 5.62 (dd, J=5.92,3.07Hz, 1H), 4.52-4.40 (m, 2H), 4.33 (d, J=5.70Hz, 1H), 4.00 (dq, J=9.98,5.01Hz, 1H), 3.75-3.61 (m, 3H), 3.35-3.07 (m, 3H),3.02-2.88(m,2H),2.38-2.23(m,1H),2.14-1.94(m,1H),1.28-1.19(m,2H),1.14-1.03 (m,2H)。

Embodiment 101

Synthesize (4S)-N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -8- methyl -7- (3- (trifluoromethyl) Phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

0 DEG C to (4S)-N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine - 4- yls) -8- methyl -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneousIn the solution of stirring of -5 (the 2H)-formamides (300mg, 0.527mmol) in methanol (10mL) be added dropwise HCl (8.00 μ L, 0.263mmol), 5min time is lasted.Then reactant mixture is stirred into 1h at 30 DEG C.(TLC eluant, eluents：5%MeOH/ DCM:R_f-0.3；UV activation) and evaporation solvent.Reactant mixture sodium bicarbonate solution is neutralized and gained solid is filtered, With ether (2x50ml), pentane (2x50ml) washing obtains pure product (4S)-N- (2- ((S) -2,3- the third oxygen of dihydroxy Base) pyridin-4-yl) -8- methyl -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (210mg, 0.380mmol, 72.1% yield), it is pale solid.LCMS(m/ z):530.29[M+H]⁺, Rt=2.04min.

¹H NMR(400MHz,CDCl₃):δ ppm 2.05 (dt, J=14.09,7.10Hz, 1H), 2.37-2.30 (m, 4H), 2.96-2.93 (m, 1H), 3.00 (dd, J=12.28,3.29Hz, 1H), 3.34-3.13 (m, 3H), 3.67-3.60 (m, 2H), 4.00-3.92 (m, 1H), 4.38 (s, 1H), 4.42 (d, J=4.60Hz, 2H), 5.63 (dd, J=5.92,3.07Hz, 1H), 6.68 (dd, J=5.81,1.86Hz, 1H), 7.03 (d, J=1.53Hz, 1H), 7.49 (s, 1H), 7.69-7.64 (m, 1H), 7.79-7.71(m,2H),7.85-7.81(m,2H),13.22(s,1H)。

Embodiment 102

Synthesize (4S) -8- chloro- 7- (1- cyclopropyl -1H- pyrazoles -4- bases)-N- (2- ((S) -2,3- dihydroxy propoxyl group) pyrroles Pyridine -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- 7- of (4S) -8- (1- cyclopropyl -1H- pyrazoles -4- bases)-N- (2- (((R) -2,2- dimethyl -1,3- Dioxolane -4- bases) methoxyl group) pyridin-4-yl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two AzepineHCl is added in the solution of stirring of -5 (the 2H)-formamides (200mg, 0.362mmol) in methanol (10mL) (0.055mL, 1.812mmol), is then stirred at room temperature 1h.(TLC systems:Rf-0.1,EtOAc).Reactant mixture is depressurized Concentrate and neutralize residue with saturated sodium bicarbonate solution.Gained solid is filtered and dried, the chloro- 7- (1- of (4S) -8- are obtained Cyclopropyl -1H- pyrazoles -4- bases)-N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -3,4- dihydro -1,4- bridges Asia Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (160mg, 0.309mmol, 85% yield), it is ash White solid.LCMS(m/z):512.00[M+H]⁺.Rt=3.15min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.81 (s, 1H), 8.10 (s, 1H), 8.02 (s, 1H) 7.98 (d, J= 5.70Hz, 1H), 7.59 (s, 1H), 7.15-6.99 (m, 2H), 5.62 (dd, J=5.92,3.07Hz (1H), 4.53-4.38 (m, 2H), 4.01 (quin, J=4.82Hz, 1H), 3.75-3.64 (m, 3H), 3.36-3.08 (m, 3H), 2.98 (dd, J=12.17, 3.18Hz, 1H), 2.32 (qd, J=9.68,4.93Hz, 1H), 2.04 (dt, J=14.25,7.34Hz, 1H), 1.29-1.17 (m,2H),1.14-1.04(m,2H)。

Embodiment 103

Synthesize (4S) -8- chloro- N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (2- methoxyl groups -5- (three Methyl fluoride) pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine

At 0 DEG C to the chloro- N- of (4S) -8- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridin-4-yl) -7- (2- methoxyl groups -5- (trifluoromethyl) pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diazaAdd in the solution of stirring of -5 (the 2H)-formamides (350mg, 0.564mmol) in methanol (5mL) Enter HCl/water solution (0.476mL, 5.64mmol), 2h is stirred at 0 DEG C.(TLC eluant, eluents：100%EtOAc:R_f-0.2；UV is activated ).Reactant mixture is alkalized (until pH-8-9) with saturated sodium bicarbonate solution and solvent is evaporated under reduced pressure.By residue water Dilution (10mL) is simultaneously extracted into dichloromethane (2x20mL).The organic extract of merging is through anhydrous sodium sulfate drying, and filtering is simultaneously Filter vacuum is evaporated, crude product is obtained.Crude compound obtains desired product (4S) -8- chloro- with triturated under ether (5mL) N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (2- methoxyl groups -5- (trifluoromethyl) pyridin-3-yl) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamide (262mg, 0.448mmol, 80% Yield), it is white solid.LCMS(m/z):581.23[M+H]⁺,R_t=2.17min.

¹H NMR(400MHz,CdCl₃):δ ppm 12.86-12.53 (m, 1H), 8.63 (d, J=1.32Hz, 1H), 7.96 (d, J=2.19Hz, 1H), 7.86 (d, J=5.92Hz, 1H), 7.66 (s, 1H), 7.00-6.97 (m, 1H), 6.62 (dd, J= 5.81,1.86Hz, 1H), 5.63 (dd, J=5.81,3.18Hz, 1H), 4.43 (d, J=5.04Hz, 2H), 4.27-4.19 (m, 1H),4.03(s,3H),3.97(br s,1H),3.72-3.58(m,2H),3.37-3.18(m,2H),3.17-3.08(m,1H), 3.02 (dd, J=12.28,3.07Hz, 1H), 2.82 (br t, J=6.47Hz, 1H), 2.40-2.26 (m, 1H), 2.08 (dt, J =14.36,7.29Hz, 1H).

Embodiment 104

Synthesize (4S) -8- cyano group-N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (3- (trifluoromethyl) Phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -8- cyano group-N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxies Base) pyridin-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two AzepineHCl/water solution is added in the solution of stirring of -5 (the 2H)-formamides (220mg, 0.379mmol) in methanol (5mL) (0.320mL, 3.79mmol) and stir 2h.(TLC eluant, eluents：100%EtOAc:R_f-0.2；UV activation).By reaction mixing Thing is alkalized (until pH-8-9) with saturated sodium bicarbonate solution and is evaporated under reduced pressure solvent.By residue diluted with water (5mL) and extract Get in dichloromethane (2x10mL).The organic extract of merging filters through anhydrous sodium sulfate drying and evaporates filter vacuum, Obtain crude product.By crude product with triturated under ether (5mL), desired product (4S) -8- cyano group-N- (2- ((S) -2,3- bis- are obtained Hydroxy propyloxy group) pyridin-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (201mg, 0.366mmol, 96% yield), it is white solid.LCMS(m/z): 541.20[M+H]⁺,R_t=2.00min.

¹H NMR(400MHz,CDCl₃):δppm 12.81-12.55(m,1H),8.17-8.01(m,2H),7.97-7.82 (m, 3H), 7.81-7.71 (m, 1H), 7.06 (d, J=1.75Hz, 1H), 6.76 (dd, J=5.81,1.86Hz, 1H), 5.69 (dd, J=5.81,2.74Hz, 1H), 4.44 (d, J=5.04Hz, 2H), 4.29-4.07 (m, 1H), 4.02-3.91 (m, 1H), 3.65(br s,2H),3.38-3.21(m,2H),3.18-3.01(m,2H),2.80(br s,1H),2.44-2.33(m,1H), 2.12 (dt, J=14.52,7.54Hz, 1H).

Embodiment 105

Synthesize (4S) -8- chloro- N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (1- methyl isophthalic acid H- pyrroles Azoles -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridin-4-yl) -7- (1- methyl isophthalic acid H- pyrazoles -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two AzepineHCl is added in the solution of stirring of -5 (the 2H)-formamides (280mg, 0.532mmol) in methanol (20mL) (2.0ml, 65.8mmol), is then stirred at room temperature 1h.(TLC systems:10%MeOH/DCM.Rf values：0.2).By reaction mixing Thing is concentrated under reduced pressure and by residue 10%NaHCO₃The aqueous solution neutralizes (30mL).The solid separated out is filtered, is dried under reduced pressure, is obtained Crude compound.Product pentane and ether (1:1) grind, obtain the chloro- N- of (4S) -8- (2- ((S) -2,3- the third oxygen of dihydroxy Base) pyridin-4-yl) -7- (1- methyl isophthalic acid H- pyrazoles -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] Diaza- 5 (2H)-formamides (150mg, 0.304mmol, 57.1% yield), it is pale solid.LCMS(m/z): 486.15[M+H]⁺, Rt=1.62min.

¹H NMR(400MHz,CDCl₃):δppm 12.85-12.83(m,1H),8.06-7.96(m,3H),7.59(s, 1H), 7.14-6.97 (m, 2H), 5.63 (dd, J=5.92,3.07Hz, 1H), 4.49-4.42 (m, 2H), 4.03 (s, 4H), 3.72-3.64 (m, 2H), 3.32-3.07 (m, 3H), 3.02-2.88 (m, 1H), 2.35-2.27 (m, 1H), 2.04 (dt, J= 14.14,7.18Hz,1H)。

Embodiment 106

Synthesize (4S) -8- chloro- N- (2- ((R) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (1- methyl isophthalic acid H- pyrroles Azoles -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (2- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridin-4-yl) -7- (1- methyl isophthalic acid H- pyrazoles -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two AzepineIn solution of -5 (the 2H)-formamides (290mg, 0.551mmol) in methanol (10mL) add HCl (2.0ml, 65.8mmol), 1h is then stirred at room temperature.Reactant mixture is evaporated under reduced pressure to remove methanol solvate, then with 10% NaHCO₃Solution (20mL) is neutralized.By gained solid filter and use water (3x20mL) fully washing, vacuum drying, obtain (4S)- The chloro- N- of 8- (2- ((R) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (1- methyl isophthalic acid H- pyrazoles -4- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(160mg, 0.327mmol, 59.3% are received -5 (2H)-formamides Rate), it is pale solid.LCMS(m/z):486.19[M+H]⁺, Rt=1.63min.

¹H NMR(400MHz,CDCl₃):δppm 12.82(s,1H),8.07-7.92(m,3H),7.59(s,1H),7.11- 7.04 (m, 2H), 5.62 (dd, J=5.92,3.07Hz, 1H), 4.49-4.40 (m, 2H), 4.36 (br d, J=4.17Hz, 1H),4.03-3.96(m,4H),3.70-3.64(m,2H),3.35-3.17(m,2H),3.14-3.09(m,1H),3.04-2.95 (m, 2H), 2.42-2.29 (m, 1H), 2.04 (dt, J=14.20,7.04Hz, 1H).

Embodiment 107

Synthesize (4S) -8- chloro- 7- (3- (difluoromethyl) phenyl)-N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridine -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- 7- of (4S) -8- (3- (difluoromethyl) phenyl)-N- (2- (((R) -2,2- dimethyl -1,3- dioxies Heterocycle pentane -4- bases) methoxyl group) pyridin-4-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaIn the solution of stirring of -5 (the 2H)-formamides (300mg, 0.524mmol) in methanol (10mL) add HCl (7.97 μ L, 0.262mmol) it is added dropwise, lasts 5min time.Then reactant mixture is stirred into 1h at 30 DEG C.(TLC eluant, eluents：5% MeOH/DCM:R_f-0.3；UV activation), and evaporation solvent.Reactant mixture is neutralized into (10mL) and mistake with sodium bicarbonate solution Filter gained solid, is washed with ether (2x50mL) and pentane (2x50ml), obtains the pure chloro- 7- of product (4S) -8- (3- (two Methyl fluoride) phenyl)-N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides (100mg, 0.188mmol, 35.8% yield), it is solid for canescence Body.LCMS(m/z):532.37[M+H]⁺, Rt=2.06min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.86 (s, 1H), 7.86 (d, J=5.48Hz, 3H), 7.73-7.57 (m, 3H), 7.01 (s, 1H), 6.91-6.58 (m, 2H), 5.65 (d, J=2.85Hz, 1H), 4.43 (d, J=4.60Hz, 2H), 4.29 (d, J=5.04Hz, 1H), 3.97 (d, J=4.82Hz, 1H), 3.73-3.53 (m, 2H), 3.37-3.18 (m, 2H), 3.17-3.10 (m, 1H), 3.02 (dd, J=12.39,2.96Hz, 1H), 2.89 (t, J=6.36Hz, 1H), 2.35 (td, J= 9.32,4.17Hz, 1H), 2.07 (dt, J=14.20,7.26Hz, 1H).

Embodiment 108

Synthesize (4S) -8- chloro- N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (2- (trifluoromethyl) pyrroles Pyridine -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides:

At 0 DEG C to the chloro- N- of (4S) -8- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridin-4-yl) -7- (2- (trifluoromethyl) pyridin-4-yl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] DiazaHCl/water is added in the solution of stirring of -5 (the 2H)-formamides (250mg, 0.423mmol) in methanol (10mL) molten Liquid (2mL, 24.00mmol), is then stirred at room temperature 1h.(TLC eluant, eluents：5%MeOH/DCM Rf-0.2；UV activation).Will Reactant mixture is concentrated under reduced pressure and by residue saturation NaHCO₃Solution neutralizes (20mL), is then extracted with 10%MeOH/DCM (2×50mL).The organic layer of merging filters through anhydrous sodium sulfate drying and filtrate evaporation is obtained into crude compound.By thick thing Through chromatogram purification, (GRACE uses C-18 reversed-phase columns, mobile phase A to matter:0.1% formic acid/water；B:Acetonitrile, eluant, eluent 55-59%B/ A).The product fraction of merging is concentrated, it is basified with saturation NaHCO3.The solid of precipitation is filtered and dried, obtain (4S)- The chloro- N- of 8- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (2- (trifluoromethyl) pyridin-4-yl) -3,4- two Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(186mg, 0.337mmol, 80% are received -5 (2H)-formamides Rate).LCMS(m/z):551.17[M+H]⁺.Rt=1.98min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 12.34 (s, 1H), 9.00 (d, J=5.04Hz, 1H), 8.27-8.31 (m, 1H), 8.14 (d, J=4.82Hz, 1H), 7.94 (s, 1H), 7.90 (d, J=5.70Hz, 1H), 7.00 (d, J=1.75Hz, 1H), 6.58 (dd, J=5.81,1.86Hz, 1H), 5.45 (dd, J=5.92,2.85Hz, 1H), 4.85 (br s, 1H), 4.58 (br d, J=1.10Hz, 1H), 4.21 (dd, J=10.74,4.60Hz, 1H), 4.10 (dd, J=10.85,6.25Hz, 1H), 3.73-3.79 (m, 1H), 3.42 (br d, J=5.26Hz, 2H), 3.23-3.28 (m, 3H), 3.06-3.17 (m, 1H), 2.22- 2.34(m,1H),1.90-2.00(m,1H)。

Embodiment 109

Synthesize (4S) -8- chloro- N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (5- (trifluoromethyl) pyrroles Pyridine -3- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides:

At 0 DEG C to the chloro- N- of (4S) -8- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridin-4-yl) -7- (5- (trifluoromethyl) pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] DiazaHCl is added in the solution of stirring of -5 (the 2H)-formamides (180mg, 0.305mmol) in methanol (10mL) (3ml, 99mmol), is then stirred at room temperature 2h.(TLC systems:10%MeOH/DCM, Rf value：0.25).Reactant mixture is used Saturation NaHCO₃Basified (until pH 8-9) is simultaneously extracted with ethyl acetate (3x30mL).The organic layer of merging is dry through sodium sulphate Dry, concentration purifies (GRACE instruments, C-18 reversed-phase columns are eluted with the formic acid of 27% acetonitrile/1%) through column chromatography, obtains (4S) -8- Chloro- N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (5- (trifluoromethyl) pyridin-3-yl) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(105mg, 0.190mmol, 62.5% are received -5 (2H)-formamides Rate), it is pale solid.LCMS(m/z):551.17[M+H]⁺.Rt=1.93min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 12.37 (s, 1H), 9.29 (d, J=1.97Hz, 1H), 9.16 (d, J =1.32Hz, 1H), 8.63 (s, 1H), 7.94 (s, 1H), 7.88 (d, J=5.70Hz, 1H), 7.00 (d, J=1.75Hz, 1H), 6.56 (dd, J=5.81,1.86Hz, 1H), 5.45 (dd, J=5.92,3.07Hz, 1H), 4.84 (d, J=5.04Hz, 1H), 4.57 (t, J=5.70Hz, 1H), 4.21 (dd, J=10.74,4.60Hz, 1H), 4.09 (dd, J=10.85,6.25Hz, 1H), 3.76 (dq, J=10.80,5.54Hz, 1H), 3.37-3.46 (m, 2H), 3.22-3.33 (m, 1H), 3.05-3.19 (m, 2H), 2.96-3.04(m,1H),2.17-2.36(m,1H),1.85-2.00(m,1H)

Embodiment 110

Synthesize (4S) -8- chloro- N- (5- ((S) -2,3- dihydroxy propoxyl group) pyridin-3-yl) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to the chloro- N- of (4S) -8- (5- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridin-3-yl) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazaHCl (5mL, 165mmol) is added in solution of -5 (the 2H)-formamides (0.4g, 0.745mmol) in methanol (6mL), is lasted 10min a period of time, 5h is then stirred at room temperature.(TLC eluant, eluents：10%MeOH/DCM, Rf value：0.2；UV activation). Reactant mixture is concentrated, residue is absorbed and (30mL) and neutralized in water with saturated sodium bicarbonate solution, water layer uses 10% MeOH/DCM extracts (3x30mL).The organic layer of merging is with salt water washing (20mL), through anhydrous sodium sulfate drying, filters and depressurizes Concentration, obtains crude compound.Thick material is merged with previous batch, 430mg is by using combiflash chromatograms altogether (silica gel is eluted with 10%MeOH/DCM) purify, obtain the chloro- N- of (4S) -8- (5- ((S) -2,3- dihydroxy propoxyl group) pyridine - 3- yls) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4]-diaza-5 (2H)-formamide (140mg, 0.272mmol, 36.5% yield), it is yellow solid.LCMS(m/z):497.35[M+H]⁺, Rt=1.32min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 12.60 (s, 1H), 8.65 (d, J=5.04Hz, 1H), 7.97 (d, J =2.63Hz, 1H), 7.93 (d, J=2.19Hz, 1H), 7.88 (s, 1H), 7.67-7.61 (m, 2H), 7.58 (dd, J=5.15, 1.21Hz, 1H), 5.46 (dd, J=5.92,3.07Hz, 1H), 4.97 (d, J=5.04Hz, 1H), 4.68 (t, J=5.59Hz, 1H), 4.02 (dd, J=9.65,3.73Hz, 1H), 3.91-3.85 (m, 1H), 3.83-3.75 (m, 1H), 3.45 (t, J= 5.70Hz,2H),3.32-3.27(m,1H),3.16-3.06(m,2H),3.02-2.96(m,1H),2.58(s,3H),2.31- 2.19(m,1H),2.01-1.91(m,1H)

Embodiment 111

Synthesize (4S) -7- chloro- N- (2- (3,3,3- trifluoro propyls) phenyl) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides

Under a nitrogen under stirring at room temperature by the chloro- 2,3,4,5- tetrahydrochysenes -1,4- endo-methylene groups pyrido [2,3- of (4S) -7- B] [1,4] diaza(250mg, 1.278mmol) is dissolved in tetrahydrofuran (THF) (10mL), sequentially adds triphosgene (190mg, 0.639mmol), TEA (0.534mL, 3.83mmol).30min is stirred at room temperature in reactant mixture.Thereto plus Enter 2- (3,3,3- trifluoro propyls) aniline (483mg, 2.56mmol) and stir 16h at 80 DEG C in seal pipe.Mix reaction Thing reaches room temperature, is gone out with 15ml water quenchings and uses 2x25ml ethyl acetate to extract, organic layer is dried over sodium sulfate and is concentrated under reduced pressure, and obtains Obtain crude compound.Purified on column chromatography is purified into (silica gel：100-200 mesh), obtain the chloro- N- of (4S) -7- (2- (3,3,3- Trifluoro propyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (240mg, 0.563mmol, 44.1% yield), it is white solid, (R_fValue：0.35,10% methanol/DCM), LCMS (m/z): 411.16[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ ppm 11.49 (s, 1H), 7.79 (dd, J=8.00,0.99Hz, 1H), 7.61 (d, J=8.11Hz, 1H), 7.35-7.31 (m, 1H), 7.24-7.29 (m, 1H), 7.17-7.09 (m, 2H), 5.44 (dd, J=5.92,3.07Hz, 1H), 3.21-2.90 (m, 6H), 2.66-2.53 (m, 2H), 2.20 (dddd, J=13.67,9.95, 6.08,3.62Hz, 1H), 1.91 (dt, J=13.70,6.96Hz, 1H).

Embodiment 112

Synthesize (4S)-N- (pyridine -2- bases) -7- (4- (trifluoromethyl) piperidin-1-yl) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 25 DEG C to (the chloro- N- of (4S) -7- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (700mg, 2.217mmol), 4- (trifluoromethyl) piperidines (679mg, 4.43mmol) In the solution of degassing in 1,4- dioxanes (20mL) add dicyclohexyl (2', 4', 6'- triisopropyl-[1,1'- biphenyl]- 2- yls) phosphine (423mg, 0.887mmol), potassium carbonate (919mg, 6.65mmol) and acid chloride (II) (100mg, 0.443mmol). Reactant mixture is stirred into 16h at 90 DEG C in seal pipe.Allow to cool to room temperature and pour the mixture into cold water (70mL) And (3x50mL) is extracted with ethyl acetate.The organic layer of merging is through anhydrous Na₂SO₄Dry and be concentrated under reduced pressure, obtain crude product.Slightly Mixture processed is through flash column chromatography (silica gel；100-200 mesh, with 1 to 2% ethanol/methylene elute), obtain (4S)- N- (pyridine -2- bases) -7- (4- (trifluoromethyl) piperidin-1-yl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (320mg, 32% yield), it is white solid (TLC:Eluant, eluent；Ethyl acetate, R_f= 0.4), LCMS (m/z):433.22[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.15 (s, 1H), 8.34-8.18 (m, 1H), 8.09 (dt, J= 8.39,0.96Hz, 1H), 7.77 (ddd, J=8.55,7.13,1.86Hz, 1H), 7.36 (d, J=8.77Hz, 1H) 7.04 (ddd, J=7.29,4.88,0.99Hz, 1H) 6.51 (d, J=8.55Hz, 1H) 5.43 (dd, J=5.92,3.07Hz, 1H) 4.39 (d, J=12.93Hz, 2H), 3.17-3.04 (m, 1H), 3.02-2.76 (m, 5H), 2.72-2.52 (m, 1H), 2.25- 2.04(m,1H),1.96-1.76(m,3H),1.57-1.39(m,2H)。

Embodiment 113

Synthesize (4S)-N- (pyridine -2- bases) -7- ((R) -3- (trifluoromethyl) piperidin-1-yl) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature to the chloro- N- of (4S) -7- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (750mg, 2.375mmol) and (70:30-R/S) -3- (trifluoromethyl) piperidines (728mg, Potassium carbonate (985mg, 7.13mmol) and acid chloride 4.75mmol) are added in the solution of the degassing in 1,4- dioxanes (20mL) (II)(107mg,0.475mmol).Reactant mixture is stirred into 16h at 90 DEG C in seal pipe.So that reactant mixture is cooled down To room temperature, pour into cold water (70mL) and be extracted with ethyl acetate (3x50mL).The organic layer of merging is through anhydrous Na₂SO₄Dry, It is concentrated under reduced pressure, obtains crude product.Crude mixture is through flash column chromatography (silica gel：100-200 mesh, with 1% to 2% methanol/ Dichloromethane eluent), obtain (4S)-N- (pyridine -2- bases) -7- ((R/S) -3- (trifluoromethyl) piperidin-1-yl) -3,4- bis- Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(the 70 of -5 (2H)-formamides:30,R:S diastereomeric) is different Structure body mixture (310mg, yield 29.6%), it is pale solid (TLC:Eluant, eluent, 100% ethyl acetate, R_f:0.4), LCMS(m/z):433.2[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.10-12.94 (m, 1H), 8.22 (d, J=5.15Hz, 1H), 8.15-7.98 (m, 1H), 7.76 (t, J=7.48Hz, 1H), 7.45-7.28 (m, 1H), 7.04 (ddd, J=7.34,4.93, 0.88Hz, 1H), 6.54 (d, J=8.55Hz, 1H), 5.48-5.32 (m, 1H), 4.35-4.12 (m, 2H), -3.12-2.89 (m, 5H),2.88-2.78(m,1H),2.61-2.52(m,1H),2.25-2.04(m,1H),2.04-1.92(m,1H),1.90-1.74 (m,2H),1.71-1.46(m,2H)。

Embodiment 114

Synthesize (4S)-N- (pyridine -2- bases) -7- ((S) -3- (trifluoromethyl) piperidin-1-yl) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides:

Room temperature to the chloro- N- of (4S) -7- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (550mg, 1.742mmol), (33:66-R/S) -3- (trifluoromethyl) piperidines (534mg, Potassium carbonate (722mg, 5.23mmol), two hexamethylenes 3.48mmol) are added in the solution of the degassing in the dioxane of Isosorbide-5-Nitrae-(20mL) Base (2', 4', 6'- triisopropyl-[1,1'- biphenyl] -2- bases) phosphine (332mg, 0.697mmol) and acid chloride (II) (78mg, 0.348mmol).Reactant mixture is stirred into 16h at 90 DEG C in seal pipe.Reactant mixture is cooled to room temperature, poured into cold In water (70mL).Crude product is extracted with ethyl acetate (3x50mL).The organic layer of merging is through anhydrous Na₂SO₄Dry, filter, decompression Concentration, obtains crude product.Crude mixture is through flash column chromatography (silica gel：100-200 mesh, with 1 to 2% methanol/dichloromethane Alkane is eluted), obtain (4S)-N- (pyridine -2- bases) -7- ((S/R) -3- (trifluoromethyl) piperidin-1-yl) -3,4- dihydro-Isosorbide-5-Nitraes - Endo-methylene group pyrido [2,3-b] [1,4] diaza(the 33 of -5 (2H)-formamides:67,S:R diastereo-isomerism mixing) Thing (250mg, 33.2% yield), it is white solid (TLC:Eluant, eluent, 100% ethyl acetate, R_f0.4), LCMS (m/z): 433.2[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.06-12.99 (m, 1H), 8.22 (d, J=5.01Hz, 1H), 8.07 (dt, J=8.33,0.88Hz, 1H), 7.76 (ddd, J=8.55,7.13,1.86Hz, 1H), 7.47-7.29 (m, 1H), 7.04 (ddd, J=7.29,4.88,0.99Hz, 1H), 6.54 (d, J=8.55Hz, 1H), 5.51-5.36 (m, 1H), 4.37- 4.15(m,2H),3.12-2.80(m,6H),2.60-2.52(m,1H),2.25-2.04(m,1H),2.04-1.93(m,1H), 1.90-1.74(m,2H),1.72-1.48(m,2H)。

Embodiment 115

Synthesize (4S)-N- (pyridine -2- bases) -7- ((R) -2- (trifluoromethyl) pyrrolidin-1-yl) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature to the chloro- N- of (4S) -7- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (700mg, 2.217mmol), (R) -2- (trifluoromethyl) pyrrolidines (617mg, 4.43mmol) in the solution of the degassing in 1,4- dioxanes (20mL) add dicyclohexyl (2', 4', 6'- triisopropyl-[1, 1'- biphenyl] -2- bases) phosphine (423mg, 0.887mmol), potassium carbonate (919mg, 6.65mmol) and acid chloride (II) (100mg, 0.443mmol).Reactant mixture is heated into 16h in seal pipe at 90 DEG C.Reactant mixture is poured into cold water (70mL) simultaneously It is extracted with ethyl acetate (3x50mL).The organic layer of merging is through anhydrous sodium sulfate drying and is concentrated under reduced pressure, and obtains crude compound. Crude mixture is through flash column chromatography (silica gel:100-200 mesh, is eluted with 1 to 2% ethanol/methylene), obtain (4S)-N- (pyridine -2- bases) -7- ((R) -2- (trifluoromethyl) pyrrolidin-1-yl) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides (220mg, 0.52mmol, 32% yield), it is pale solid (TLC:100% ethyl acetate, R_f=0.4), LCMS (m/z):419.21[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.16 (s, 1H), 8.20 (d, J=6.33,1H), 8.06 (d, J= 8.33Hz, 1H), 7.77 (td, J=7.84,1.86Hz, 1H), 7.40 (d, J=8.55Hz, 1H), 7.05 (ddd, J=7.29, 4.88,0.99Hz, 1H), 6.36 (d, J=8.77Hz, 1H), 5.40 (dd, J=5.92,3.07Hz, 1H), 5.28-5.06 (m, 1H), 3.84 (dt, J=10.14,5.12Hz, 1H), 3.65-3.36 (m, 1H), 3.17-2.91 (m, 2H), 2.90-2.80 (m, 2H), 2.25-2.04 (m, 5H), 1.84 (dt, J=13.65,7.10Hz, 1H).

Embodiment 116

Synthesize (4S)-N- (pyridine -2- bases) -7- (2- (trifluoromethyl) morpholino) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature to the chloro- N- of (4S) -7- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (700mg, 2.217mmol), 2- (trifluoromethyl) morpholine (688mg, 4.43mmol) 1, Dicyclohexyl (2', 4', 6'- triisopropyl-[1,1'- biphenyl] -2- is added in the solution of degassing in 4- dioxanes (20mL) Base) phosphine (423mg, 0.887mmol), potassium carbonate (919mg, 6.65mmol) and acid chloride (II) (100mg, 0.443mmol).Will Reactant mixture heats 16h in seal pipe at 90 DEG C, is subsequently cooled to room temperature, is subsequently poured into cold water (70mL) and uses acetic acid Ethyl ester extracts (3x50mL).The organic layer of merging is through anhydrous sodium sulfate drying and is concentrated under reduced pressure, and obtains crude product.Crude mixture Through flash column chromatography (silica gel：100-200 mesh, with 1 to 2% ethanol/methylene elute), obtain (4S)-N- (pyridine- 2- yls) -7- (2- (trifluoromethyl) morpholino) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (300mg, 0.69mmol, 46% yield), it is pale solid (TLC:100% ethyl acetate, R_f= 0.4), LCMS (m/z):435.20[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ ppm 12.99 (d, J=4.82Hz, 1H), 8.20 (dt, J=4.82, 0.88Hz, 1H), 8.07 (dd, J=8.44,0.77Hz, 1H), 7.77 (td, J=7.84,1.86Hz, 1H), 7.43 (d, J= 8.55Hz, 1H), 7.05 (ddd, J=7.29,4.88,0.77Hz, 1H), 6.62 (d, J=8.55Hz, 1H), 5.44 (ddd, J= 9.98,6.25,3.29Hz, 1H), 4.34 (dd, J=7.13,3.40Hz, 1H), 4.25 (d, J=12.28Hz, 1H), 4.14 (dd, J=11.62,1.75Hz, 1H), 4.10-3.94 (m, 1H), 3.87-3.67 (m, 1H), 3.15-2.91 (m, 6H), 2.30- 2.14(m,1H),1.72-1.65(m,1H)。

Embodiment 117

Synthesis (4S)-N- (4- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

0 DEG C to (4S)-N- (4- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine - 2- yls) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 It is added dropwise in the solution of stirring of (the 2H)-formamide (450mg, 0.810mmol) in dichloromethane (12mL) and water (1.12mL) 4.0M hydrochloric acid dioxane solutions (1.68mL, 0.810mmol), last 5min time.Then by reactant mixture at 30 DEG C Stir 3h and evaporation solvent.Reactant mixture sodium bicarbonate solution is neutralized and (3 × 50mL) is extracted with ethyl acetate, then By organic layer water, the aqueous salt solu-tion of merging, and it is dried over sodium sulfate, and evaporated, thick material is obtained, it is brown solid (TLC eluant, eluents：5%MeOH/DCM:R_f=0.2；UV activation).Crude compound is washed with pentane, obtain pure (4S)- N- (4- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- bridges are sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (340mg, 0.635mmol, 78.0% yield), it is White solid, LCMS (m/z):516.3[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.42 (s, 1H) 8.54-8.44 (m, 2H) 8.10 (d, J= 5.92Hz, 1H) 7.85-7.81 (m, 1H) 7.80-7.68 (m, 4H) 6.71 (dd, J=5.70,2.41Hz, 1H) 5.51 (dd, J= 5.92,3.07Hz, 1H) 5.01 (d, J=5.26Hz, 1H) 4.70 (t, J=5.70Hz, 1H) 4.11 (dd, J=9.76, 3.84Hz, 1H) 3.96 (dd, J=9.87,6.36Hz, 1H) 3.87-3.79 (m, 1H) 3.51-3.42 (m, 2H) 3.26-3.17 (m, 1H) 3.15-3.06 (m, 2H) 2.96 (dd, J=12.06,3.29Hz, 1H) 2.25 (dddd, J=13.65,9.92,6.14, 3.62Hz, 1H) 1.95 (dt, J=13.81,6.91Hz, 1H).

Embodiment 118

Synthesis (4S)-N- (4- ((R) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S)-N- (4- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) under nitrogen gas stirring Methoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (500mg, 0.900mmol) are molten in dichloromethane (12.5mL) and water (1.25ml) It is disposable in liquid to add 4.0M hydrochloric acid dioxane solutions (2mL, 0.900mmol), last 1min.By reactant mixture at 30 DEG C Stir 3h and evaporation solvent.Reactant mixture sodium bicarbonate solution is neutralized and is extracted with ethyl acetate (3 × 50mL).Merge Organic layer water, aqueous salt solu-tion, and through anhydrous sodium sulfate drying and evaporate, obtain thick material, it is brown solid (TLC eluant, eluents：5%MeOH/DCM:R_f=0.2；UV activation).Crude compound is washed with pentane, obtain pure (4S)- N- (4- ((R) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- bridges are sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (yield of 340mg, 0.612mmol 68.0%), it is White solid, LCMS (m/z):516.2[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.42 (s, 1H) 8.58-8.44 (m, 2H) 8.10 (d, J= 5.70Hz, 1H) 7.85-7-68 (m, 5H) 6.71 (dd, J=5.81,2.30Hz, 1H) 5.52 (dd, J=5.70,3.07Hz, 1H) 5.01 (d, J=5.26Hz, 1H) 4.70 (t, J=5.59Hz, 1H) 4.11 (dd, J=9.76,3.84Hz, 1H) 3.96 (dd, J= 9.76,6.25Hz,1H)3.87-3.79(m,1H)3.50-3.43(m,2H)3.27-3.17(m,1H)3.16-3.05(m,2H) 2.96 (dd, J=12.06,3.07Hz, 1H) 2.30-2.20 (m, 1H) 1.95 (dt, J=13.92,7.07Hz, 1H).

Embodiment 119

Synthesize (4S)-N- (2- (((R)-tetrahydrofuran -3- bases) epoxide) pyrimidine-4-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

30 DEG C to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diazaTriethylamine is added in the solution of the stirring of (600mg, 1.965mmol) in THF (20mL) (0.822mL, 5.90mmol) and triphosgene (292mg, 0.983mmol), then in mutually synthermal stirring 1h.Then add at 30 DEG C Enter (R) -2- ((tetrahydrofuran -3- bases) epoxide) pyrimidine -4- amine (1068mg, 5.90mmol) and add reactant mixture at 70 DEG C Hot 16h.Reactant mixture is reached room temperature and solvent is evaporated under reduced pressure, by the residue diluted with water (15mL) of acquisition and use DCM Extract (2x20ml).The organic layer water of merging, salt water washing, and through anhydrous sodium sulfate drying.Organic solvent is evaporated under reduced pressure, obtains To crude product.Crude mixture purifies (formic acid/water and acetonitrile 30%) through preparation HPLC, obtains (4S)-N- (2- (((R)-four Hydrogen furans -3- bases) epoxide) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (310mg, 0.605mmol, 30.6% yield), it is pale solid. (TLC:R_f=0.25,10%MeOH/EtOAc), LCMS (m/z):513.26[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.61 (s, 1H), 8.50 (d, J=7.49Hz, 1H), 8.45 (d, J =5.62Hz, 1H), 8.25 (s, 1H), 7.87 (d, J=7.67Hz, 1H), 7.81-7.68 (m, 4H), 5.47 (dd, J=5.92, 3.07Hz, 1H), 5.32 (ddt, J=6.49,4.36,2.14,2.14Hz, 1H), 3.91-3.76 (m, 1H), 3.76-3.59 (m, 3H), 3.28-3.05 (m, 3H), 2.96 (dd, J=12.06,3.29Hz, 1H), 2.38-2.15 (m, 1H), 2.14-1.88 (m, 3H)。

Embodiment 120

Synthesize (4S)-N- (pyridine -2- bases) -7- ((S) -2- (trifluoromethyl) pyrrolidin-1-yl) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

25 DEG C to the chloro- N- of (4S) -7- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (700mg, 2.217mmol), (S) -2- (trifluoromethyl) pyrrolidines (617mg, 4.43mmol) in the solution of the degassing in 1,4- dioxanes (20mL) add dicyclohexyl (2', 4', 6'- triisopropyl-[1, 1'- biphenyl] -2- bases) phosphine (423mg, 0.887mmol) and acid chloride (II) (100mg, 0.443mmol).Reactant mixture is existed In seal pipe 16h is heated at 90 DEG C.Reactant mixture is poured into cold water (70mL) and is extracted with ethyl acetate (3x50mL).Close And organic layer is through anhydrous sodium sulfate drying and is concentrated under reduced pressure, obtain crude product.Crude mixture is through flash column chromatography (silicon Glue:100-200 mesh, with 1 to 2%MeOH/DCM elution), obtain (4S)-N- (pyridine -2- bases) -7- ((S) -2- (trifluoromethyl) Pyrrolidin-1-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (250mg, 0.598mmol, 35% yield), it is white solid (TLC:100% ethyl acetate, R_f=0.4), LCMS (m/z): 419.18[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.13 (s, 1H), 8.30-8.18 (d, J=7.2,1H), 8.18- 7.96 (d, J=8.40,1H), 7.77 (ddd, J=8.55,7.13,1.86Hz, 1H), 7.41 (d, J=8.55Hz, 1H), 7.04 (ddd, J=7.23,4.82,1.10Hz, 1H), 6.38 (d, J=8.55Hz, 1H), 5.48 (dd, J=6.03,3.18Hz, 1H), 5.12-5.01 (m, 1H), 3.84 (m, 1H), 3.49 (q, J=9.65, J=5.70Hz, 1H), 3.04-2.90 (m, 4H), 2.33- 2.12(m,5H),1.72-1.80(m,1H)。

Embodiment 121

Synthesize (4S)-N- (2,3- dihydrobenzos [b] [1,4] Dioxin -6- bases) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaIn the solution of (500mg, 1.982mmol) in THF (30ml) add triphosgene (294mg, 0.991mmol), 1h is then stirred at room temperature.Then it is sequentially added 2,3- dihydrobenzos [b] [1,4] Dioxin -6- Amine (899mg, 5.94mmol) and triethylamine (1.381mL, 9.91mmol).Reactant mixture is heated in seal pipe at 70 DEG C 16h.Reactant mixture is poured into saturation NaHCO₃In solution (150mL) and it is extracted with ethyl acetate (3x50mL).What is merged has Machine layer is through anhydrous Na₂SO₄Dry and be concentrated under reduced pressure, obtain crude compound.Crude mixture is through flash column chromatography (silica gel: 100-200 mesh, the gradient mixture using 1% ethanol/methylene is used as eluant, eluent), obtain (4S)-N- (2,3- dihydrobenzenes And [b] [1,4] Dioxin -6- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides (315mg, 0.734mmol, 51% yield), it is brown solid (TLC:5% ethanol/methylene, R_f=0.3), LCMS (m/z):430.0[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ ppm 12.69 (s, 1H), 8.59 (d, J=5.26Hz, 1H), 7.75-7.59 (m, 4H), 7.16 (s, 1H), 6.96-6.75 (dd, J=5.81,2.41,2H), 5.46 (dd, J=5.81,3.18Hz, 1H), 4.27-4.10(m,4H),3.24-3.02(m,2H),3.00-2.76(m,2H),2.50(s,3H),2.25-2.09(m,1H), 1.91-1.71(m,1H)。

Embodiment 122

Synthesize (4S)-N- (6- ethyl pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaIn the solution of (500mg, 1.982mmol) in THF (25ml) add triphosgene (294mg, 0.991mmol), then stir to room temperature, continue 1h.Then 6- ethyl pyrazine -2- amine (317mg, 2.58mmol) is sequentially added, Triethylamine (1.381mL, 9.91mmol) and in seal pipe in 70 DEG C of heating response mixture 16h.Reactant mixture is poured into Saturation NaHCO₃In solution (150mL) and it is extracted with ethyl acetate (3x50mL).The organic layer of merging is through anhydrous Na₂SO₄Dry simultaneously It is concentrated under reduced pressure, obtains crude product.Crude mixture purifies (silica gel through column chromatography：100-200 mesh, eluant, eluent：1% methanol/dichloro Methane), obtain (4S)-N- (6- ethyl pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydros-Isosorbide-5-Nitrae-endo-methylene group Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (260mg, 0.640mmol, 32.3% yield), it is yellowish Color solid (TLC:5%MeOH/DCM, R_f=0.3), LCMS (m/z):402.23[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.47 (s, 1H), 9.23 (s, 1H), 8.59 (d, J=5.26Hz, 1H), 8.29 (s, 1H), 8.02 (d, J=4.82Hz, 2H), 7.88 (s, 1H), 7.64-7.84 (d, J=7.68Hz, 1H), 5.52 (dd, J=6.03,2.96Hz, 1H), 3.08-3.33 (m, 2H), 2.89-3.08 (m, 2H), 2.78 (q, J=7.60Hz, 2H), 2.57(s,3H),2.39-2.13(m,1H),2.12–1.88(m,1H)1.21-1.30(m,3H)。

Embodiment 123

Synthesis (4S)-N- (6- ((S) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

0 DEG C to (4S)-N- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine - 2- yls) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (600mg, 1.078mmol) is in CH₂Cl₂The two of 4N HCl is added dropwise in solution in (16mL) and water (1.6mL) Oxane solution (2.24mL, 8.96mmol), lasts 5min time.Then 3h is stirred at room temperature in reactant mixture.Will reaction Mixture is concentrated under reduced pressure, and obtains crude product.Crude product with water dilutes and with sodium acid carbonate (15ml) neutralization and by water layer EtOAc (2x15ml) is extracted.The organic layer water of merging, salt water washing, and through anhydrous Na₂SO₄Dry, organic solvent is evaporated under reduced pressure, obtains To crude product.Crude product triturated under ether, obtains (4S)-N- (6- ((S) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine (346mg, 64% yield, 0.67mmol), it is pale solid (TLC:5%MeOH/CH₂Cl₂,R_f=0.3), LCMS (m/ z):517.18[M+H]⁺。

¹H NMR(400MHz,DMSO-d6):δ 13.14 (s, 1H), 8.95 (s, 1H), 8.36 (d, J=7.7Hz, 1H), 8.22 (s, 1H), 7.99 (s, 1H), 7.89-7.79 (m, 2H), 7.74 (q, J=8.0Hz, 2H), 5.54 (s, 1H), 4.95 (d, J =5.1Hz, 1H), 4.64 (t, J=5.5Hz, 1H), 4.09 (qd, J=10.7,5.1Hz, 2H), 3.77 (d, J=5.2Hz, 1H), 3.45-3.33 (m, 2H), 3.23-3.06 (m, 3H), 2.97 (dd, J=12.0,3.3Hz, 1H), 2.25 (ddt, J= 14.0,9.8,4.8Hz, 1H), 1.97 (dt, J=14.1,7.3Hz, 1H).

Embodiment 124

(4S) -7- ((R) -2- methyl morpholine generations)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (700mg, 2.210mmol), (R) -2- methyl morpholine hydrochlorides (335mg, 2.431mmol)、Cs₂CO₃(1440mg, 4.42mmol) and 2- dicyclohexyl phosphino- -2', 4 ', 6 '-tri isopropyl biphenyl (421mg, 0.884mmol) adds Pd (OAc) in 1,4- dioxanes (10mL) in the solution of degassing₂(99mg, 0.442mmol).Then reactant mixture is heated into 16h at 100 DEG C.So that reactant mixture reaches room temperature, 2x15ml water quenchings are used Go out and use 2x50ml ethyl acetate to extract.The organic layer of merging is through anhydrous sodium sulfate drying and is concentrated under reduced pressure, and obtains rough chemical combination Thing.Purified on column chromatography is purified, and obtains (4S) -7- ((R) -2- methyl morpholine generations)-N- (pyrazine -2- bases) -3,4- dihydro -1, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(275mg, 0.696mmol, 31.5% are received -5 (2H)-formamides Rate), it is pale solid (TLC:R_fValue：0.3,5% methanol/DCM), LCMS (m/z):383.28[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ 13.40 (s, 1H), 9.38 (d, J=1.3Hz, 1H), 8.46-8.17 (m, 2H), 7.40 (d, J=8.6Hz, 1H), 6.53 (d, J=8.6Hz, 1H), 5.44 (dd, J=6.1,3.1Hz, 1H), 4.20- 4.04(m,1H),4.00-3.93(m,1H),3.91-3.80(m,1H),3.70-3.53(m,2H),3.17-3.05(m,1H), 3.03-2.92 (m, 2H), 2.87 (ddd, J=19.7,11.9,3.4Hz, 2H), 2.58 (dd, J=12.6,10.4Hz, 1H), 2.21-2.12 (m, 1H), 1.86 (dd, J=14.3,7.2Hz, 1H), 1.22 (d, J=6.2Hz, 3H).

Embodiment 125

Synthesize (4S) -7- ((S) -2- methyl morpholine generations)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (700mg, 2.210mmol), (S) -2- methyl morpholine hydrochlorides (335mg, 2.431mmol)、Cs₂CO₃(1440mg, 4.42mmol) and 2- dicyclohexyl phosphino- -2', 4 ', 6 '-tri isopropyl biphenyl (421mg, 0.884mmol) adds Pd (OAc) in 1,4- dioxanes (10mL) in the solution of degassing₂(99mg, 0.442mmol).Then reactant mixture is heated into 16h at 100 DEG C.So that reactant mixture reaches room temperature, 2x15ml water quenchings are used Go out and use 2x50ml ethyl acetate to extract.The organic layer of merging is through anhydrous sodium sulfate drying and is concentrated under reduced pressure, and obtains rough chemical combination Thing.Purified on column chromatography is purified, and obtains (4S) -7- ((S) -2- methyl morpholine generations)-N- (pyrazine -2- bases) -3,4- dihydro -1, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(285mg, 0.725mmol, 32.8% are received -5 (2H)-formamides Rate), it is pale solid (TLC:R_fValue：0.3,5% methanol/DCM), LCMS (m/z):383.28[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ 13.40 (s, 1H), 9.38 (d, J=1.4Hz, 1H), 8.40-8.14 (m, 2H), 7.40 (d, J=8.6Hz, 1H), 6.53 (d, J=8.6Hz, 1H), 5.44 (dd, J=6.0,3.1Hz, 1H), 4.12 (s, 1H), 3.96 (ddd, J=11.5,3.6,1.4Hz, 1H), 3.86 (d, J=12.8Hz, 1H), 3.62 (dd, J=11.8, 2.8Hz, 2H), 3.09 (dd, J=11.4,8.4Hz, 1H), 2.96 (s, 2H), 2.93-2.78 (m, 2H), 2.58 (dd, J= 12.6,10.4Hz, 1H), 2.17 (s, 1H), 1.85 (dt, J=14.3,7.3Hz, 1H), 1.23 (d, J=6.2Hz, 3H).

Embodiment 126

Synthesize (4S)-N- (3,4- dihydro -2H- benzos [b] [1,4] dioxas- 7- bases) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaIn the solution of (500mg, 1.982mmol) in THF (30ml) add triphosgene (294mg, 0.991mmol), then stir to room temperature, continue 1h.Then it is sequentially added 3,4- dihydro -2H- benzos [b] [1,4] dioxa - 7- amine (426mg, 2.58mmol) and triethylamine (1.381mL, 9.91mmol) are simultaneously mixed in seal pipe in 70 DEG C of heating responses Thing 16h.Reactant mixture is poured into saturation NaHCO₃Solution (150mL) is simultaneously extracted with ethyl acetate (3x50mL).What is merged has Machine layer is through anhydrous Na₂SO₄Dry and be concentrated under reduced pressure, obtain crude product.Crude mixture is through flash column chromatography (silica gel；100- 200 mesh, the gradient mixture using 1%MeOH/DCM is used as eluant, eluent), obtain (4S)-N- (3,4- dihydro -2H- benzos [b] [1,4] dioxa- 7- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (394mg, 0.889mmol, 68% yield), it is faint yellow solid (TLC:5%MeOH/ DCM,R_f=0.3), LCMS (m/z):444.28[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ ppm 12.76 (s, 1H), 8.59 (d, J=5.04Hz, 1H), 7.79-7.58 (m, 3H), 7.24-7.06 (m, 2H), 6.96 (d, J=8.77Hz, 1H), 5.46 (dd, J=5.70,3.07Hz, 1H), 4.10 (dt, J=19.51,5.37Hz, 4H), 3.15-3.02 (m, 1H), 3.02-2.77 (m, 3H), 2.50 (s, 3H), 2.46-2.18 (m,1H),2.17-2.01(m,2H),2.01-1.83-(m,2H)。

Embodiment 127

Synthesis (4S)-N- (2- ((R) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

0 DEG C to (4S)-N- (2- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine - 4- yls) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 4.0M hydrochloric acid is added in solution of (the 2H)-formamide (450mg, 0.809mmol) in dichloromethane (12mL), water (1.2mL) In 1,4- dioxanes (1.68mL, 0.809mmol).Reactant mixture is stirred into 3h and concentrated solvent at 30 DEG C.Reaction is mixed Compound is distributed between water (10mL) and EtOAc (25mL).Organic layer water, aqueous salt solu-tion, through anhydrous Na₂SO₄Dry ＆ Filter and evaporate filtrate, obtain thick material (TLC eluant, eluents：5%MeOH/DCM:R_f-0.2；UV activation).Thick material is pure Change, washed and be suspended in water (5mL) with pentane, stir 15min and filter, obtain pure (4S)-N- (2- ((R) -2,3- Dihydroxy propoxyl group) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza- 5 (2H)-formamides (260mg, 0.480mmol, 59.4% yield), it is pale solid, LCMS (m/z):517.2[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ 13.58 (s, 1H), 8.60 (d, J=7.5Hz, 1H), 8.46 (d, J= 5.6Hz, 1H), 8.27 (s, 1H), 7.91-7.78 (m, 3H), 7.75-7.60 (m, 2H), 5.48 (dd, J=5.9,3.0Hz, 1H), 4.91 (d, J=4.8Hz, 1H), 4.62 (t, J=5.5Hz, 1H), 4.22 (dd, J=10.8,4.4Hz, 1H), 4.13 (dd, J=10.7,5.9Hz, 1H), 3.80 (q, J=5.4Hz, 1H), 3.43 (t, J=5.7Hz, 2H), 3.21 (s, 1H), 3.16-3.05 (m, 2H), 2.96 (dd, J=12.0,3.3Hz, 1H), 2.29-2.10 (m, 1H), 2.04-1.84 (m, 1H).

Embodiment 128

Synthesis (4S)-N- (6- ((R) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

0 DEG C to (4S)-N- (6- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine - 2- yls) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 4.0M hydrochloric acid is added in solution of (the 2H)-formamide (300mg, 0.539mmol) in dichloromethane (8.0mL), water (0.8mL) In the dioxane of Isosorbide-5-Nitrae-(1.12mL, 0.539mmol), 1min (dropwise addition) is lasted.Reactant mixture is stirred into 3h at 30 DEG C and dense Contracting solvent.By reactant mixture distribution between water (10mL) and ethyl acetate (25mL).Organic layer is washed with water, saline solution Wash, through anhydrous Na₂SO₄Dry and evaporate, obtain crude compound (TLC eluant, eluents：5%MeOH/DCM:R_f=0.2；UV is activated ).Crude compound is washed with pentane and is suspended in water (5mL), is stirred 15min and is filtered, obtains pure (4S)-N- (6- ((R) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (280mg, 0.533mmol, 99% yield), it is greyish white Color solid, LCMS (m/z):517.3[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ 13.15 (s, 1H), 8.95 (s, 1H), 8.36 (d, J=7.7Hz, 1H), 8.22 (s, 1H), 7.99 (s, 1H), 7.86 (t, J=7.8Hz, 1H), 7.80 (d, J=7.8Hz, 1H), 7.74 (q, J= 8.0Hz, 2H), 5.52 (dd, J=5.8,3.0Hz, 1H), 4.95 (d, J=5.1Hz, 1H), 4.64 (t, J=5.5Hz, 1H), 4.14 (dd, J=10.7,4.0Hz, 1H), 4.03 (dd, J=10.7,6.1Hz, 1H), 3.77 (q, J=5.2Hz, 1H), 3.36 (hept, J=5.8Hz, 2H), 3.22 (s, 1H), 3.12 (d, J=11.3Hz, 2H), 2.97 (dd, J=11.9,3.3Hz, 1H), 2.26 (t, J=7.0Hz, 1H), 2.03-1.90 (m, 1H).

Embodiment 129

Synthesize (4S)-N- (6- ethyoxyl pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature by DMAP (726mg, 5.94mmol) and (6- ethyoxyl pyrazine -2- bases) phenyl carbamates (1541mg, 5.94mmol) adds to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diazaIn the solution of the stirring of (500mg, 1.982mmol) in THF.By mixture in seal pipe 80 DEG C are heated to, 20h is kept.It is cooled to after room temperature, evaporation solvent and crude product are through flash column chromatography (silica gel：100-200 Mesh, 5%MeOH/DCM is used as eluant, eluent).By the material absorbing of recovery in ethanol, the solid isolated is divided through filtering From obtaining (4S)-N- (6- ethyoxyl pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro-Isosorbide-5-Nitrae-endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (416mg, 0.978mmol, 49.3% yield), it is canescence Solid (TLC eluant, eluents：10%MeOH/DCM, R_f:0.4), LCMS (m/z):418.28[M+H]⁺。

¹H NMR(DMSO-d6,400MHz):δ 13.18 (s, 1H), 8.95 (s, 1H), 8.53 (dd, J=5.0,1.0Hz, 1H), 7.98 (d, J=0.5Hz, 1H), 7.86-7.79 (m, 2H), 7.73 (s, 2H), 5.51 (dd, J=5.9,3.1Hz, 1H), 4.22 (qd, J=7.1,2.3Hz, 2H), 3.25-3.17 (m, 1H), 3.11 (dt, J=11.7,2.4Hz, 2H), 2.97 (dd, J =12.0,3.3Hz, 1H), 2.58 (s, 3H), 2.25 (dddd, J=13.7,9.9,6.2,3.8Hz, 1H), 2.04-1.90 (m, 1H), 1.27 (t, J=7.0Hz, 3H).

Embodiment 130

Synthesis (4S) -7- (2- picoline -4- bases)-N- (1H- pyrazolos [3,4-c] pyridine -5- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaIn the solution of the stirring of (300mg, 1.189mmol) in THF (10mL) add triphosgene (176mg, 0.594mmol) and stirring 30min.After 30 minutes, triethylamine (0.829mL, 5.94mmol) and 1H- pyrazolos [3,4-c] are added Pyridin-5-amine (191mg, 1.427mmol).Reactant mixture is stirred into 16h in seal pipe at 65-70 DEG C.Reactant mixture Gone out with 2x15ml water quenchings and use 2x15ml ethyl acetate to extract.The organic layer of merging is through anhydrous sodium sulfate drying and is concentrated under reduced pressure, Obtain crude compound.Crude mixture is purified through column chromatography, obtains (4S) -7- (2- picoline -4- bases)-N- (1H- pyrazoles And [3,4-c] pyridine -5- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first Acid amides (330mg, 0.764mmol, 64.3% yield), it is faint yellow solid (R_fValue：0.25, net ethyl acetate), LCMS (m/ z):413.24[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ 13.47 (s, 2H), 8.89 (t, J=1.1Hz, 1H), 8.71-8.57 (m, 1H), 8.44 (t, J=1.0Hz, 1H), 8.28-8.10 (m, 2H), 8.05-7.93 (m, 1H), 7.79 (d, J=8.0Hz, 1H), 7.71 (d, J=8.0Hz, 1H), 5.56 (dd, J=5.9,3.1Hz, 1H), 3.22 (tt, J=8.1,3.0Hz, 1H), 3.15- 3.04 (m, 2H), 2.97 (dd, J=12.0,3.3Hz, 1H), 2.67 (s, 3H), 2.36-2.16 (m, 1H), 1.96 (dd, J= 14.0,7.1Hz,1H)。

Embodiment 131

Synthesize (4S) -7- (2- picoline -4- bases)-N- (quinoxaline -5- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaIn the solution of (500mg, 1.982mmol) in THF (30ml) add triphosgene (294mg, 0.991mmol), 1h is then stirred at room temperature.Then quinoxaline -5- amine (374mg, 2.58mmol) and three are added in room temperature in succession Ethamine (1.381mL, 9.91mmol), then heats 16h by reactant mixture at 70 DEG C.Reactant mixture is poured into saturation NaHCO₃In solution (150mL) and it is extracted with ethyl acetate (3x50mL).The organic layer of merging is through anhydrous Na₂SO₄Dry and depressurize Concentration, obtains crude product.Crude product is through flash column chromatography (silica gel：100-200 mesh, is mixed using 1%MeOH/DCM gradient Compound is used as eluant, eluent), obtain (4S) -7- (2- picoline -4- bases)-N- (quinoxaline -5- bases) -3,4- dihydros-Isosorbide-5-Nitrae-bridge Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (300mg, 0.709mmol, 55% yield), it is Faint yellow solid (TLC:5%MeOH/DCM, R_f=0.3), LCMS (m/z):424.26[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.50 (s, 1H), 8.91 (s, 1H), 8.68 (dd, J=7.89, 1.10Hz, 1H), 8.48 (d, J=5.26Hz, 1H), 7.92-7.80 (m, 2H), 7.80-7.62 (m, 4H), 5.58 (dd, J= 5.81,2.96Hz,1H),3.20-3.08(m,4H),3.08-2.86(m,1H),2.26(s,4H),2.00-1.95(m,1H)。

Embodiment 132

Synthesis (4S)-N- (2- ((R) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (3- (trifluoromethyl) phenyl) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

0 DEG C to (4S)-N- (2- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine - 4- yls) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 4.0M hydrochloric acid is added dropwise in solution of (the 2H)-formamide (450mg, 0.810mmol) in dichloromethane (12mL), water (1.2mL) Solution in the solution (1.68mL, 0.810mmol) of the dioxane of Isosorbide-5-Nitrae-, continues 1min.By reactant mixture in 30 DEG C of stirrings 3h and concentrated solvent.By reactant mixture distribution between water (10mL) and EtOAc (25mL).Organic layer water, saline solution Washing, through anhydrous Na₂SO₄Dry and evaporate, obtain crude compound (TLC eluant, eluents：5%MeOH/DCM:R_f-0.2；UV is activated ).Crude compound is washed with pentane, and compound is suspended in water (5mL), is stirred 15min and is filtered, obtains pure (4S)-N- (2- ((R) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (310mg, 0.600mmol, 74.0% yield), It is white solid, LCMS (m/z):516.3[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ 13.02 (s, 1H), 8.27-8.15 (m, 2H), 7.94 (d, J=5.7Hz, 1H), 7.89 (d, J=7.8Hz, 1H), 7.82 (t, J=7.7Hz, 1H), 7.72 (d, J=8.0Hz, 1H), 7.66 (d, J= 8.0Hz, 1H), 7.11 (d, J=1.8Hz, 1H), 6.80 (dd, J=5.8,1.9Hz, 1H), 5.47 (dd, J=5.9,3.0Hz, 1H), 4.85 (d, J=5.1Hz, 1H), 4.59 (t, J=5.7Hz, 1H), 4.23 (dd, J=10.9,4.6Hz, 1H), 4.13 (dd, J=10.9,6.2Hz, 1H), 3.81-3.65 (m, 1H), 3.43 (t, J=5.6,1.3Hz, 2H), 3.25-3.03 (m, 3H), 2.97 (dd, J=12.0,3.3Hz, 1H), 2.25 (m, J=13.8,9.7,4.7Hz, 1H), 1.95 (m, J=14.3, 7.6Hz,1H)。

Embodiment 133

(4S) -7- (2- picoline -4- bases)-N- (1H- pyrazolos [4,3-b] pyridine -5- bases) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaIn the solution of the stirring of (600mg, 2.378mmol) in THF (10mL) add triphosgene (353mg, 1.189mmol) and stirring 30min, thereto add triethylamine (1.657mL, 11.89mmol) and 1H- pyrazolos [4,3-b] pyrrole Pyridine -5- amine (319mg, 2.378mmol).Reactant mixture is stirred into 16h at 60 DEG C.Reactant mixture 2x15ml water quenchings are gone out simultaneously Extracted with 2x15ml ethyl acetate.The organic layer of merging is through anhydrous sodium sulfate drying and is concentrated under reduced pressure, and obtains crude compound.Slightly Product purifies (100-200 mesh) through column chromatography, obtains (4S) -7- (2- picoline -4- bases)-N- (1H- pyrazolos [4,3-b] Pyridine -5- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (265mg, 0.622mmol, 26.2% yield), it is faint yellow solid (TLC:R_fValue：0.25, net ethyl acetate), LCMS (m/ z):413.28[M+H]⁺。

¹H NMR(400MHz,DMSO-d6):δ 13.55 (s, 1H), 13.27 (s, 1H), 8.66 (dd, J=5.4,0.8Hz, 1H), 8.32 (d, J=9.2Hz, 1H), 8.16-7.94 (m, 4H), 7.94-7.51 (m, 2H), 5.55 (dd, J=6.0,3.1Hz, 1H), 3.21 (d, J=8.7Hz, 1H), 3.18-3.05 (m, 2H), 2.97 (dd, J=12.0,3.3Hz, 1H), 2.69 (s, 3H), 2.35-2.17 (m, 1H), 1.96 (dt, J=14.2,7.6Hz, 1H).

Embodiment 134

Synthesis (4S)-N- (2- ((S) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

0 DEG C to (4S)-N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine - 4- yls) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 4.0M hydrochloric acid is added dropwise in solution of (the 2H)-formamide (350mg, 0.629mmol) in dichloromethane (8mL), water (0.8mL) The dioxane solution of Isosorbide-5-Nitrae-(1.12mL, 0.629mmol), lasts 1min.Reactant mixture is stirred into 3h and concentrated solvent at 30 DEG C. By reactant mixture distribution between water (10mL) and EtOAc (25mL).Organic layer water, aqueous salt solu-tion, through anhydrous Na₂SO₄Dry and evaporate, obtain crude compound (TLC eluant, eluents：5%MeOH/DCM:R_f=0.2；UV activation).By thick thing Matter is purified by chiral separation, obtains (4S)-N- (2- ((S) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (3- (trifluoros Methyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (210mg, 0.405mmol, 64.3% yield), it is pale solid, LCMS (m/z):517.3[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ 13.59 (s, 1H), 8.60 (d, J=7.5Hz, 1H), 8.46 (d, J= 5.7Hz, 1H), 8.27 (s, 1H), 7.90-7.78 (m, 3H), 7.76-7.69 (m, 2H), 5.48 (dd, J=5.8,3.0Hz, 1H), 4.91 (d, J=5.0Hz, 1H), 4.63 (t, J=5.5Hz, 1H), 4.46-4.01 (m, 2H), 3.80 (h, J=5.5Hz, 1H), 3.44 (t, J=5.5Hz, 2H), 3.21 (s, 1H), 3.12 (dd, J=10.1,5.5Hz, 2H), 2.96 (dd, J=12.0, 3.3Hz, 1H), 2.24 (d, J=8.7Hz, 1H), 2.03-1.90 (m, 1H).

Embodiment 135

Synthesize (4S) -7- (2,6- lutidines -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

28 DEG C by tripotassium phosphate (804mg, 3.79mmol) add to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (600mg, 1.894mmol), (2,6- bis- Picoline -4- bases) stirring of the boric acid (343mg, 2.273mmol) in n-butyl alcohol (10mL) and water (1.667mL) solution In.Reactant mixture is deaerated 15min, Pd is added₂(dba)₃(87mg, 0.095mmol), and X-phos (90mg, 0.189mmol).Reactant mixture is deaerated 15min again, and reactant mixture is stirred into 3hr at 100 DEG C.By reactant mixture 28 DEG C are cooled to, is then distributed between water (10mL) and EtOAc (25mL).Organic layer is separated, then through anhydrous Na₂SO₄It is dry It is dry, filter and evaporate filtrate, obtain thick material, it is brown solid (TLC eluant, eluents：10%MeOH/EtOAc；R_f=0.3； UV activation).Thick material is purified through column chromatography, is used (100-200 mesh) silica gel and is eluted with 5-10%MeOH/EtOAc, is obtained Pure (4S) -7- (2,6- lutidines -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza- 5 (2H)-formamides (355mg, 0.916mmol, 48.3% yield), it is faint yellow solid, LCMS(m/z):387.44[M+H]⁺。

¹H NMR(DMSO-d₆,400MHz,):δ 13.77 (s, 1H), 9.43 (d, J=1.5Hz, 1H), 8.53-8.28 (m, 2H), 7.91-7.59 (m, 4H), 5.51 (dd, J=6.0,3.1Hz, 1H), 3.25-2.81 (m, 4H), 2.56-2.46 (m, 6H), 2.26 (dddd, J=13.7,9.9,6.1,3.7Hz, 1H), 1.97 (dq, J=14.4,7.5,6.8Hz, 1H).

Embodiment 136

Synthesize (4S) -7- (2- cyclopropyl pyridine -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

28 DEG C by tripotassium phosphate (670mg, 3.16mmol) add to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (500mg, 1.579mmol), (2- rings third Yl pyridines -4- bases) stirring of the boric acid (309mg, 1.894mmol) in 1,4- dioxanes (10mL) and water (1.667mL) it is molten In liquid.Reactant mixture is deaerated 15min, Pd is added₂(dba)₃(72.3mg, 0.079mmol), and X-phos (75mg, 0.158mmol).Reactant mixture is deaerated 15min again, and reactant mixture is stirred into 3hr at 100 DEG C.By reactant mixture 28 DEG C are cooled to, is then distributed between water (10mL) and EtOAc (25mL).Organic layer is separated, then through anhydrous Na₂SO₄It is dry It is dry, filter and evaporate filtrate, obtain thick material, it is brown solid (TLC eluant, eluents：10%MeOH/EtOAc；R_f-0.3； UV activation).Thick material is purified through column chromatography, is used (100-200 mesh) silica gel and is eluted with 5-10%MeOH/EtOAc, is obtained Pure (4S) -7- (2- cyclopropyl pyridine -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza- 5 (2H)-formamides (260mg, 0.645mmol, 40.8% yield), it is pale solid, LCMS

(m/z):400.28[M+H]⁺。

¹H NMR(DMSO-d₆,400MHz):δ 13.75 (s, 1H), 9.43 (d, J=1.5Hz, 1H), 8.54 (d, J= 5.4Hz, 1H), 8.46-8.28 (m, 2H), 8.19-8.01 (m, 1H), 7.89-7.62 (m, 3H), 5.52 (dd, J=6.0, 3.1Hz, 1H), 3.30-2.86 (m, 4H), 2.27 (ddd, J=13.2,6.4,4.1Hz, 2H), 1.98 (dt, J=14.5, 7.6Hz,1H),1.17-0.88(m,4H).

Embodiment 137

Synthesize (4S) -7- (2- isopropyl pyridine -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

28 DEG C by tripotassium phosphate (670mg, 3.16mmol) add to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (500mg, 1.579mmol), (2- isopropyls Yl pyridines -4- bases) stirring of the boric acid (313mg, 1.894mmol) in 1,4- dioxanes (10mL) and water (1.667mL) it is molten In liquid.Reactant mixture is deaerated 15min, Pd is added₂(dba)₃(72.3mg, 0.079mmol) and X-phos (75mg, 0.158mmol).Reactant mixture is deaerated 15min again, and reactant mixture is stirred into 3hr at 100 DEG C.By reactant mixture 28 DEG C are cooled to, is then distributed between water (10mL) and EtOAc (25mL).Organic layer is separated, then through anhydrous Na₂SO₄It is dry It is dry, filter and evaporate filtrate, obtain thick material, it is brown solid (TLC eluant, eluents：10%MeOH/EtOAc:R_f=0.3； UV activation).Thick material is purified through column chromatography, uses (100-200 mesh) silica gel, and is eluted with 5-10%MeOH/EtOAc, Obtain pure (4S) -7- (2- isopropyl pyridine -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (320mg, 0.793mmol, 50.2% yield), it is pale solid. LCMS(m/z):402.33[M+H]⁺。

¹H NMR(CDCl_3,400MHz):δ 13.65 (s, 1H), 9.57 (d, J=1.5Hz, 1H), 8.68 (d, J=5.2Hz, 1H), 8.44-8.07 (m, 2H), 8.02-7.87 (m, 1H), 7.75-7.58 (m, 2H), 7.48 (d, J=8.0Hz, 1H), 5.73 (dd, J=6.0,3.2Hz, 1H), 3.43-3.14 (m, 4H), 3.04 (dd, J=12.1,3.3Hz, 1H), 2.35 (dt, J= 11.7,5.0Hz, 1H), 2.22-2.00 (m, 1H), 1.41 (dd, J=6.9,0.9Hz, 6H)

Embodiment 138

Synthesis (4S)-N- (1- methyl isophthalic acid H-1,2,3- triazole-4-yls) -7- (2- picoline -4- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Triphosgene (0.588g, 1.982mmol) is added into (4S) -7- (2- picoline -4- bases) -2,3,4,5- at 0 DEG C Tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(0.5g, 1.982mmol) is in tetrahydrofuran (20mL) Stirring solution in, then add triethylamine at 0 DEG C and (0.829mL, 5.94mmol) and stir 30min.Addition 1- methyl- 1H-1,2,3- triazole -4- amine hydro-chloride (0.4g, 2.97mmol), then heat 48h at 70 DEG C.By reaction mixing Thing is cooled to 28 DEG C, then distributes between water (10mL) and ethyl acetate (50mL).Separate organic layer and washed with water and salt Wash.Organic layer is through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain rough yellow solid (TLC eluant, eluents：10% MeOH/EtOAc:R_f=0.2；UV activation).Crude compound is washed through 100-200 silica gel purifications with 5%MeOH/ ethyl acetate It is de-, obtain (4S)-N- (1- methyl isophthalic acids H-1,2,3- triazole-4-yls) -7- (2- picoline -4- bases) -3,4- dihydros-Isosorbide-5-Nitrae-bridge Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (0.260g, 0.686mmol, 34.6% yield), its For pale solid, LCMS (m/z):377.28[M+H]⁺。

¹H NMR(400MHz,DMSO-d6):δ 13.52 (s, 1H), 8.58 (dd, J=5.3,0.8Hz, 1H), 8.07 (s, 1H), 8.00 (d, J=1.7Hz, 1H), 7.85-7.75 (m, 2H), 7.71 (d, J=8.0Hz, 1H), 5.46 (dd, J=5.9, 3.0Hz, 1H), 4.06 (s, 3H), 3.30 (s, 1H), 3.24-3.05 (m, 2H), 2.96 (dd, J=12.0,3.2Hz, 1H), 2.63 (s, 3H), 2.23 (dd, J=13.7,9.8,6.1,3.7Hz, 1H), 2.00-1.86 (m, 1H).

Embodiment 139

Synthesize (4S) -7- (2- ethylpyridine -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

Tripotassium phosphate (804mg, 3.79mmol) is added into the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (600mg, 1.894mmol), (2- ethylpyridines -4- Base) stirring of the boric acid (343mg, 2.273mmol) in n-butanol (10mL) and water (1.667mL) solution in.Reaction is mixed Compound degassing 15min.Add Pd₂(dba)₃(87mg, 0.095mmol) and X-phos (90mg, 0.189mmol).By reaction mixing Thing deaerates 15min again, then stirs 5hr at 100 DEG C.Reactant mixture is cooled to 28 DEG C, then distribute in water (10mL) and Between EtOAc (25mL).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filtering.Evaporation filtrate obtains thick material, and (TLC is washed De- agent：10%MeOH/EtOAc:R_f:0.2；UV activation).Crude compound passes through (100-200 mesh) silica gel purification, with 20% MeOH/EtOAc is eluted, and obtains (4S) -7- (2- ethylpyridine -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro-Isosorbide-5-Nitrae-bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (418mg, 1.074mmol, 56.7% yield), it is ash White solid, LCMS (m/z):388.26[M+H]⁺。

¹H NMR(DMSO-d₆,400MHz):δ 13.72 (s, 1H), 9.43 (d, J=1.5Hz, 1H), 8.62 (dd, J= 5.3,0.8Hz, 1H), 8.53-8.21 (m, 2H), 8.08 (d, J=1.7Hz, 1H), 7.97-7.64 (m, 3H), 5.52 (dd, J= 6.0,3.1Hz,1H),3.30(s,1H),3.23-3.08(m,3H),3.03-2.81(m,2H),2.34-2.16(m,1H),1.99 (d, J=6.8Hz, 1H), 1.31 (t, J=7.6Hz, 3H).

Embodiment 140

Synthesize (4S) -7- (2- ethoxy pyridine -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Tripotassium phosphate (804mg, 3.79mmol) is added into the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (600mg, 1.894mmol), (2- ethoxy pyridines- 4- yls) stirring of the boric acid (380mg, 2.273mmol) in n-butyl alcohol (10mL) and water (1.667mL) solution in.Will reaction Mixture degassing 15min, adds Pd₂(dba)₃(87mg, 0.095mmol), and X-phos (90mg, 0.189mmol).Will reaction Mixture deaerates 15min again, then stirs 4hr at 100 DEG C.Reactant mixture is cooled to 28 DEG C, then distributed in water Between (100mL) and EtOAc (120mL).EtOAc layers of separation, then through anhydrous Na₂SO₄Dry, filter and evaporate filtrate To thick material (TLC eluant, eluents：10%MeOH/EtOAc:R_f=0.4；UV activation).Crude compound passes through (100-200 mesh) silicon Glue purification, with 10%MeOH/ ethyl acetate elute, obtain (4S) -7- (2- ethoxy pyridine -4- bases)-N- (pyrazine -2- bases) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (435mg, 1.063mmol, 56.1% yield), it is pale solid, LCMS (m/z):404.28[M+H]⁺。

¹H NMR(DMSO-d₆,400MHz):δ 13.73 (s, 1H), 9.39 (d, J=1.5Hz, 1H), 8.50-8.33 (m, 1H), 8.31-8.21 (m, 2H), 7.79 (d, J=8.0Hz, 1H), 7.77-7.62 (m, 2H), 7.60 (d, J=2.2Hz, 1H), 5.50 (dd, J=5.9,3.0Hz, 1H), 4.37 (q, J=7.0Hz, 2H), 3.30 (s, 1H), 3.25-3.03 (m, 2H), 2.97 (dd, J=12.1,3.3Hz, 1H), 2.36-2.12 (m, 1H), 2.07-1.87 (m, 1H), 1.38 (t, J=7.0Hz, 3H).

Embodiment 141

Synthesize (4S) -7- (2- methoxypyridine -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Tripotassium phosphate (804mg, 3.79mmol) is added into the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (600mg, 1.894mmol), (2- methoxypyridines- 4- yls) stirring of the boric acid (348mg, 2.273mmol) in 1,4- dioxanes (12mL) and water (2.000mL) solution in.Will Reactant mixture degassing 15min, adds Pd₂(dba)₃(87mg, 0.095mmol) and X-phos (90mg, 0.189mmol).Will be anti- Answer mixture to deaerate again 15min, then stir 5hr at 100 DEG C.Reactant mixture is cooled to 28 DEG C, then distributed in water Between (25mL) and EtOAc (60mL).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained thick Material (TLC eluant, eluents：10%MeOH/EtOAc；R_f=0.2；UV activation).Crude compound is pure through (100-200 mesh) silica gel Change, eluted with 20%MeOH/ ethyl acetate, obtain (4S) -7- (2- methoxypyridine -4- bases)-N- (pyrazine -2- bases) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (310mg, 0.793mmol, 41.8% yield), it is faint yellow solid, LCMS (m/z):390.20[M+H]⁺。

¹H NMR(DMSO-d₆, 400MHz):δ 13.71 (s, 1H), 9.38 (d, J=1.5Hz, 1H), 8.43 (dd, J= 2.5,1.5Hz, 1H), 8.38 (s, 1H), 8.37 (d, J=2.5Hz, 1H), 7.75 (d, J=22.0Hz, 1H), 7.77-7.62 (m, 2H), 7.60 (s, 1H), 5.50 (dd, J=5.9,3.0Hz, 1H), 3.94 (s, 3H), 3.30 (s, 1H), 3.25-3.03 (m, 2H), 3.03-2.88 (m, 1H), 2.25 (dd, J=9.9,3.9Hz, 1H), 1.98 (dd, J=14.2,7.2Hz, 1H).

Embodiment 142

Synthesize (4S) -7- (2- methoxyl group -6- picoline -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Tripotassium phosphate (804mg, 3.79mmol) is added into the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (600mg, 1.894mmol), (2- methoxyl group -6- first Yl pyridines -4- bases) stirring of the boric acid (380mg, 2.273mmol) in 1,4- dioxanes (12mL) and water (2.000mL) it is molten In liquid.Reactant mixture is deaerated 15min, Pd is added₂(dba)₃(87mg, 0.095mmol) and X-phos (90mg, 0.189mmol).Reactant mixture is deaerated 15min again, then 5hr are stirred at 100 DEG C.Reactant mixture is cooled to 28 DEG C, Then distribute between water (25mL) and EtOAc (60mL).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filtering.Evaporation Filtrate obtains thick material (TLC eluant, eluents：10%MeOH/EtOAc；R_f=0.3；UV activation).Crude compound passes through (100- 200 mesh) silica gel purification, with 20%MeOH/ ethyl acetate elute, obtain (4S) -7- (2- methoxyl group -6- picoline -4- bases) - N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (225mg, 0.547mmol, 28.9% yield), it is pale solid, LCMS (m/z):404.24[M+H]⁺。

¹H NMR(DMSO-d₆,400MHz):δ 13.72 (s, 1H), 9.41 (d, J=1.5Hz, 1H), 8.55-8.26 (m, 2H), 7.86-7.64 (m, 3H), 7.31 (dd, J=1.4,0.7Hz, 1H), 5.51 (dd, J=6.0,3.1Hz, 1H), 3.91 (s, 3H), 3.22-3.00 (m, 3H), 2.97 (dd, J=12.1,3.3Hz, 1H), 2.63-2.41 (m, 3H), 2.25 (dddd, J= 13.8,9.8,6.1,3.6Hz, 1H), 1.97 (dt, J=14.3,7.6Hz, 1H).

Embodiment 143

Synthesis (4S)-N- (1- methyl isophthalic acid H-1,2,4- triazole -3- bases) -7- (2- picoline -4- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Triphosgene (0.588g, 1.982mmol) is added into (4S) -7- (2- picoline -4- bases) -2,3,4,5- at 0 DEG C Tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(0.5g, 1.982mmol) is in tetrahydrofuran (20mL) Stirring solution in and stirring 30min.Triethylamine (0.829mL, 5.94mmol) and 1- methyl isophthalic acids H-1,2,4- are added at 0 DEG C Triazole -3- amine (0.486g, 4.95mmol).Reactant mixture is stirred into 48h at 70 DEG C.Reactant mixture is cooled to 28 DEG C, Then distribute between water (50mL) and DCM (50mL).The organic layer water and salt water washing of separation.Organic layer is through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain rough yellow solid (TLC eluant, eluents：10%MeOH/EtOAc:R_f- 0.3；UV activation).Crude compound passes through (100-200 mesh) silica gel purification, and is eluted with 5%MeOH/ ethyl acetate, obtains (4S)-N- (1- methyl isophthalic acid H-1,2,4- triazole -3- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (0.215g, 0.563mmol, 28.4% yield), it is pale yellow Color solid, LCMS (m/z):377.28[M+H]⁺。

¹H NMR(400MHz,DMSO-d6):δ 13.44 (s, 1H), 8.55 (d, J=5.2Hz, 1H), 8.36 (s, 1H), 7.96 (s, 1H), 7.86-7.61 (m, 3H), 5.44 (dd, J=5.8,3.0Hz, 1H), 3.83 (s, 3H), 3.27 (d, J= 30.5Hz, 3H), 2.95 (dd, J=12.0,3.2Hz, 1H), 2.59 (s, 3H), 2.23 (ddt, J=16.9,9.7,4.4Hz, 1H), 1.91 (dt, J=14.7,7.4Hz, 1H).

Embodiment 144

Synthesize (4S) -7- (2- (methylamino) pyridin-4-yl)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (500mg, 1.579mmol), N- methyl -4- (4,4,5,5- tetramethyl -1,3,2- dioxies The miscellaneous amyl- 2- yls of boron heterocycle) pyridine -2- amine (443mg, 1.894mmol) and tripotassium phosphate (670mg, 3.16mmol) be in 1,4- bis- Pd (OAc) is added in oxane (15mL) and water (2mL) in the solution of degassing₂(17.72mg, 0.079mmol) and dicyclohexyl (2', 4', 6'- triisopropyl-[1,1'- biphenyl] -2- bases) phosphine (75mg, 0.158mmol) simultaneously adds reactant mixture at 120 DEG C Hot 4h.N-butanol solvent is evaporated under reduced pressure, the residue diluted with water of acquisition is simultaneously extracted (2x50mL) with DCM.The organic layer of merging With water, salt water washing, through anhydrous sodium sulfate drying and solvent is evaporated under reduced pressure, obtains crude product.Crude product is pure through flash column chromatography Change (silica gel：100-200 mesh, eluant, eluent：3%MeOH/DCM), (4S) -7- (2- (methylamino) pyridin-4-yl)-N- is obtained (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (325mg, 0.820mmol, 51.9% yield), it is white solid (TLC:10%MeOH/EtOAc, R_f:0.2), LCMS (m/ z):389.2[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.64 (s, 1H), 9.40 (d, J=1.53Hz, 1H), 8.46-8.31 (m, 2H), 8.14 (d, J=5.48Hz, 1H), 7.71 (m, J=8.11Hz, 1H), 7.61 (d, J=8.11Hz, 1H), 7.22 (dd, J=5.37,1.64Hz, 1H), 7.03 (d, J=0.88Hz, 1H), 6.40 (q, J=4.75Hz, 1H), 5.51 (dd, J= 5.92,3.07Hz, 1H), 3.35-3.04 (m, 3H), 2.96 (dd, J=12.06,3.29Hz, 1H), 2.85 (d, J=5.04Hz, 3H), 2.25 (dddd, J=13.59,9.87,6.03,3.62Hz, 1H), 1.97 (dt, J=13.87,6.99Hz, 1H).

Embodiment 145

(4S)-N- (1H- imidazos [4,5-c] pyridin-4-yl) -7- (2- picoline -4- bases) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature by (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- bridge methylenes under nitrogen gas stirring Yl pyridines simultaneously [2,3-b] [1,4] diaza(700mg, 2.77mmol) is dissolved in tetrahydrofuran (THF) (10mL), adds three Phosgene (412mg, 1.387mmol), is then stirred at room temperature 30min, and triethylamine (1.933mL, 13.87mmol) is added thereto With 1H- imidazos [4,5-c] pyridine -4- amine (372mg, 2.77mmol), reactant mixture is then stirred into 16hr at 60 DEG C.Instead Answer mixture 2x15ml water quenchings to go out and use 2x15ml ethyl acetate to extract, organic layer is dried over sodium sulfate and is concentrated under reduced pressure, and obtains To crude compound.Purified on column chromatography purifies (100-200 mesh), obtain pure compound (4S)-N- (1H- imidazos [4, 5-c] pyridin-4-yl) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine- 5 (2H)-formamides (225mg, 0.531mmol, 19.14% yield), it is faint yellow solid.(R_fValue：0.4, 10% methanol/DCM), LCMS (m/z):413.1[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 14.03 (s, 1H), 11.93 (s, 1H), 8.65 (d, J=5.3Hz, 1H), 8.35-7.86 (m, 3H), 7.86-7.37 (m, 4H), 5.72 (dd, J=5.9,3.2Hz, 1H), 3.21 (m, 3H), 3.06 (dd, J =12.2,3.3Hz, 1H), 2.78 (s, 3H), 2.38 (dddd, J=14.0,9.9,6.2,4.2Hz, 1H), 2.13 (dt, J= 14.3,7.2Hz,1H)。

Embodiment 146

Synthesize (4S) -7- (2- methoxy pyrimidine -5- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature by potassium phosphate (402mg, 1.894mmol) and (2- methoxy pyrimidine -5- bases) boric acid (175mg, 1.137mmol) add to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] DiazaStirring of -5 (the 2H)-formamides (300mg, 0.947mmol) in n-butyl alcohol (6mL) and water (2.0mL) mixture Solution in, by mixture argon-degassed 25min, then add Pd₂(dba)₃(43.4mg, 0.047mmol) and X-phos (45.2mg, 0.095mmol), 2h is heated at 120 DEG C.Reactant mixture is reached room temperature, (40mL) is diluted with water and acetic acid is used Ethyl ester extracts (3x30mL), with salt water washing (30mL).The organic layer concentration of separation, through flash column chromatography (silica gel：100- 200 mesh, 80% ethyl acetate/petroleum ether is used as eluant, eluent).The substance migration ethanol and pentane of recovery are recrystallized, obtained (4S) -7- (2- methoxy pyrimidine -5- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (190mg, 0.481mmol, 50.8% yield), it is pale solid.(flowing Phase:100% ethyl acetate, R_f:0.1), LCMS (m/z):391.2[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 13.64 (s, 1H), 9.52 (d, J=1.5Hz, 1H), 9.24 (s, 2H), 8.40- 8.25 (m, 2H), 7.64 (d, J=7.9Hz, 1H), 7.34 (d, J=8.0Hz, 1H), 5.70 (dd, J=6.0,3.1Hz, 1H), 4.12 (s, 3H), 3.33-3.13 (m, 3H), 3.02 (d, J=3.3Hz, 1H), 2.36 (dddd, J=13.9,9.9,5.9, 4.0Hz,1H),2.17-2.04(m,1H)。

Embodiment 147

Synthesize (4S) -7- (6- (hydroxymethyl) pyridin-3-yl)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature by potassium phosphate (535mg, 2.53mmol) and (6- (hydroxymethyl) pyridin-3-yl) boric acid (232mg, 1.515mmol) add to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] Diaza- 5 (2H)-formamide (400mg, 1.263mmol) stirring in the mixture of n-butyl alcohol (6mL) and water (2.0mL) In the solution mixed.By mixture argon-degassed 10min, Pd is then added₂(dba)₃(57.8mg, 0.063mmol) and X-phos (60.2mg,0.126mmol).Mixture is heated into 20h at 120 DEG C.Reactant mixture is reached room temperature, and pass through Celite pad Filtering, (10mL × 2) are washed with ethyl acetate, the solvent of filtrate is removed under reduced pressure, crude compound is obtained.Above-mentioned rough thing is through fast Fast column chromatography purifies (silica gel:100-200 mesh, is eluted with 80% ethyl acetate/n-hexane), obtain (4S) -7- (6- (hydroxyl first Base) pyridin-3-yl)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (225mg, 0.558mmol, 44.2% yield), it is pale solid.(TLC:Eluant, eluent：100% acetic acid Ethyl ester R_f:0.1), LCMS (m/z):390.35(M+H)⁺。

¹HNMR(400MHz,CDCl₃):δ 13.73 (s, 1H), 9.54 (d, J=1.3Hz, 1H), 9.19 (dd, J=2.3, 0.9Hz, 1H), 8.51 (dd, J=8.2,2.3Hz, 1H), 8.44-8.20 (m, 2H), 7.64 (d, J=8.0Hz, 1H), 7.50- 7.39 (m, 2H), 5.71 (dd, J=6.0,3.2Hz, 1H), 4.86 (d, J=2.4Hz, 2H), 3.67 (s, 1H), 3.46-3.13 (m, 3H), 3.03 (dd, J=12.1,3.3Hz, 1H), 2.43-2.26 (m, 1H), 2.19-1.98 (m, 1H).

Embodiment 148

Synthesize (4S)-N- (2,3- dihydros-[1,4] Dioxin simultaneously [2,3-b] pyridin-7-yl) -7- (2- methyl Pyridin-4-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature by (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- bridge methylenes under nitrogen gas stirring Yl pyridines simultaneously [2,3-b] [1,4] diaza(700mg, 2.77mmol) is dissolved in tetrahydrofuran (THF) (10mL), is added Simultaneously 30min is stirred at room temperature in triphosgene (412mg, 1.387mmol), and triethylamine (1.933mL, 13.87mmol) is added thereto With 2,3- dihydros-[1,4] Dioxin simultaneously [2,3-b] pyridine -7- amine (422mg, 2.77mmol).By reactant mixture 16hr is stirred at 60 DEG C.Reactant mixture is gone out with 2x15ml water quenchings and uses 2x20ml ethyl acetate to extract, and organic layer is through sodium sulphate Dry and be concentrated under reduced pressure, obtain crude compound (800mg).Purified on column chromatography is purified, and obtains pure compound (4S)-N- (2,3- dihydros-[1,4] Dioxin simultaneously [2,3-b] pyridin-7-yl) -7- (2- picoline -4- bases) -3,4- two Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamide (325mg, 0.732mmol, 26.4% Yield), it is faint yellow solid.R_fValue：0.4,10% methanol/DCM, LCMS (m/z):413.28[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ 12.81 (s, 1H), 8.58 (d, J=5.2Hz, 1H), 7.82 (d, J= 2.4Hz, 1H), 7.78-7.40 (m, 5H), 5.45 (dd, J=5.9,3.0Hz, 1H), 4.46-4.31 (m, 2H), 4.29-4.16 (m, 2H), 3.19 (d, J=8.8Hz, 1H), 3.14-3.03 (m, 2H), 2.96 (dd, J=12.0,3.3Hz, 1H), 2.55 (s, 3H), 2.23 (d, J=7.0Hz, 1H), 1.93 (dd, J=14.1,7.1Hz, 1H).

Embodiment 149

Synthesize (4R) -7- (2- picoline -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the chloro- N- of (4R) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (700mg, 2.210mmol), (2- picoline -4- bases) boric acid (454mg, 3.31mmol) and K₃PO₄X-Phos is added in the solution of the degassing of (1825mg, 6.63mmol) in 1,4- dioxanes (20mL) and water (5.00mL) (211mg, 0.442mmol) and Pd₂(dba)₃(202mg,0.221mmol).Reactant mixture is heated into 2h at 110 DEG C, makes its cold But to room temperature.(100mL) is diluted with water and is extracted with ethyl acetate (3x100mL).The organic layer of merging is through anhydrous Na₂SO₄Dry And be concentrated under reduced pressure, obtain crude product.Crude product obtains (4R) -7- (2- picoline -4- bases)-N- through flash column chromatography (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (392mg, 49.2% yield), it is pale solid (TLC:Eluant, eluent；10% ethanol/methylene, R_f=0.3), LCMS (m/z):374.18[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.76 (s, 1H), 9.43 (d, J=1.53Hz, 1H), 8.59 (d, J =5.26Hz, 1H), 8.46 (dd, J=2.63,1.53Hz, 1H), 8.39 (d, J=2.41Hz, 1H), 8.31 (s, 1H), 8.16- 8.06 (m, 1H), 7.92-7.78 (m, 1H), 7.78-7.69 (m, 1H), 5.52 (dd, J=5.81,3.18Hz, 1H), 3.37 (m, 1H), 3.28-3.06 (m, 2H), 2.98 (dd, J=11.95,3.18Hz, 1H), 2.61 (s, 3H), 2.41-2.15 (m, 1H), 2.05-1.84(m,1H)。

Embodiment 150

((4S) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-yls) (morpholino) ketone

In room temperature to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- bridge methylenes under nitrogen gas stirring Yl pyridines simultaneously [2,3-b] [1,4] diazaThe solution of (700mg, 2.77mmol) in tetrahydrofuran (THF) (10mL) is added Simultaneously 30min is stirred at room temperature in triphosgene (412mg, 1.387mmol), and triethylamine (1.933mL, 13.87mmol) is added thereto With morpholine (314mg, 3.61mmol).Reactant mixture is stirred into 16hr at 60 DEG C.Reactant mixture 2x15ml water quenchings are gone out simultaneously Extracted with 2x20ml ethyl acetate, organic layer is dried over sodium sulfate and is concentrated under reduced pressure, and obtains crude compound.Crude product is added to 100-200 silicagel columns are simultaneously eluted with 4% methanol/DCM, obtain pure compound ((4S) -7- (2- picoline -4- bases) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-yls) (morpholino) ketone (270mg, 0.735mmol, 26.5% yield), it is Light brown solid (R_fValue：0.4,10% methanol/DCM), LCMS (m/z):366.19 [M+H]⁺。

¹H NMR (400MHz, DMSO-d6) δ 8.52 (d, J=5.2Hz, 1H), 7.85-7.75 (m, 1H), 7.70 (dd, J =5.2,1.7Hz, 1H), 7.57-7.30 (m, 1H), 4.38 (s, 1H), 3.50 (d, J=72.2Hz, 8H), 3.23-2.95 (m, 1H), 2.89 (dd, J=11.6,3.2Hz, 1H), 2.58-2.32 (m, 6H), 2.33-1.92 (m, 2H)

Embodiment 151

((4S) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-yls) (piperidin-1-yl) ketone

In room temperature to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- bridge methylenes under nitrogen gas stirring Yl pyridines simultaneously [2,3-b] [1,4] diazaAdd in the solution of (700mg, 2.77mmol) in tetrahydrofuran (THF) (10mL) Enter triphosgene (412mg, 1.387mmol), 30min is then stirred at room temperature.Thereto add triethylamine (1.933mL, 13.87mmol) with piperidines (236mg, 2.77mmol).Reactant mixture is stirred into 16hr at 60 DEG C.Reactant mixture is used 2x15ml water quenchings are gone out and use 2x20ml ethyl acetate to extract, and organic layer is dried over sodium sulfate and is concentrated under reduced pressure, and obtains rough chemical combination Thing.Purified on column chromatography is purified, and obtains pure compound ((4S) -7- (2- picoline -4- bases) -3,4- dihydros-Isosorbide-5-Nitrae-bridge Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-yls) (piperidin-1-yl) ketone (220mg, 0.583mmol, 21.03% yield), it is brown solid (R_fValue：0.4,10% methanol/DCM), LCMS (m/z):364.21[M+H]⁺。

¹H NMR (400MHz, DMSO-d6) δ 8.50 (d, J=5.2Hz, 1H), 7.79 (d, J=1.8Hz, 1H), 7.74- 7.62 (m, 1H), 7.51-7.30 (m, 2H), 4.33 (t, J=4.2Hz, 1H), 3.49 (d, J=71.1Hz, 4H), 3.22-2.95 (m, 2H), 2.88 (dd, J=11.6,3.2Hz, 1H), 2.61-2.35 (m, 6H), 2.24-1.94 (m, 1H), 1.55 (s, 5H).

Embodiment 152

Synthesize (4S) -7- (2- picoline -4- bases)-N- (pyrido [3,4-b] pyrazine -5- bases) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaIn the solution of (600mg, 2.378mmol) in THF (30ml) add triphosgene (353mg, 1.189mmol), then stir to room temperature, continue 1h.Then pyrido [3,4-b] pyrazine -5- amine is added in room temperature in succession (452mg, 3.09mmol) and triethylamine (1.657mL, 11.89mmol), in 70 DEG C of heating response mixtures in seal pipe 16h.Reactant mixture is poured into saturation NaHCO₃In solution (150mL) and it is extracted with ethyl acetate (250mL).What is merged is organic Layer is through anhydrous Na₂SO₄Dry and be concentrated under reduced pressure, obtain crude product.Crude mixture is through flash column chromatography (silica gel:100- 200 mesh, the gradient mixture using 1%MeOH/DCM is used as eluant, eluent), obtain (4S) -7- (2- picoline -4- bases)-N- (pyrido [3,4-b] pyrazine -5- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (260mg, 0.613mmol, 38% yield), it is brown solid (TLC:5%MeOH/DCM, R_f=0.3), LCMS(m/z):425.27[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.84 (s, 1H), 9.11 (d, J=1.97Hz, 1H), 8.58 (d, J =5.70Hz, 1H), 8.37-8.51 (m, 1H), 8.19 (d, J=1.97Hz, 1H), 7.59-7.78 (m, 5H), 5.54-5.50 (m, 1H), 3.22-3.33 (m, 1H), 3.07-3.19 (m, 1H), 3.00 (dd, J=11.95,3.18Hz, 1H), 2.50 (dt, J =3.67,1.78Hz, 1H), 2.21-2.33 (m, 4H), 1.98 (dt, J=13.92,7.07Hz, 1H).

Embodiment 153

Synthesize (4S)-N- (3- methylcinnolin -5- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaIn the solution of (600mg, 2.378mmol) in THF (30ml) add triphosgene (353mg, 1.189mmol), then stir to room temperature, continue 1h.Then in succession room temperature add 3- methylcinnolin -5- amine (492mg, 3.09mmol) with triethylamine (1.657mL, 11.89mmol), reactant mixture is heated into 16h at 70 DEG C.Reactant mixture is fallen Enter saturation NaHCO₃In solution (150mL) and it is extracted with ethyl acetate (2x50mL).The organic layer of merging is through anhydrous Na₂SO₄Dry And be concentrated under reduced pressure, obtain crude product.Crude mixture is through flash column chromatography (silica gel:100-200 mesh, uses 1%MeOH/ DCM gradient mixture is used as eluant, eluent), obtain (4S)-N- (3- methylcinnolin -5- bases) -7- (2- picoline -4- bases) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (280mg, 0.647mmol, 42% yield), it is faint yellow solid (TLC:5%MeOH/DCM, R_f=0.25), LCMS (m/z):438.26[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.43 (s, 1H), 9.11 (s, 1H), 8.49 (d, J=5.92Hz, 1H), 8.39 (d, J=7.45Hz, 1H), 8.19 (s, 1H), 7.81 (t, J=8.11Hz, 1H), 7.65 (d, J=7.5Hz 1H), 7.35-7.31 (m, 3H), 5.77 (dd, J=5.81,3.18Hz, 1H), 3.41-3.15 (m, 3H), 2.91 (m, 1H), 2.30- 2.25(m,3H),2.24(s,3H),2.10-1.92(m,1H)。

Embodiment 154

Synthesize (4S) -7- (5- picoline -3- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines with argon-degassed 20min And [2,3-b] [1,4] diaza- 5 (2H)-formamides (600mg, 1.894mmol), (5- picoline -3- bases) boric acid (311mg, 2.273mmol) and K₃PO₄(804mg, 3.79mmol) is molten in 1,4- dioxanes (12.00mL), water (4.00mL) X-Phos (90mg, 0.189mmol) and three (dibenzalacetone) two palladium (0) (87mg, 0.095mmol) is added in liquid, again Use argon-degassed 10min.Reactant mixture is stirred into 4h at 100 DEG C, room temperature is subsequently cooled to.By reactant mixture through diatomite Filter, then filtrate water dilutes and extracted (3 × 10mL) with EtOAc.Organic layer water, the aqueous salt solu-tion of merging, and It is dried over sodium sulfate, and evaporated, obtain crude compound (TLC eluant, eluents：5%MeOH/DCM:R_f-0.3；UV activation).It is rough Compound is purified through column chromatography, using 100-200 silica gel mesh and with 1 to 3%MeOH/DCM elution, obtains pure (4S) -7- (5- Picoline -3- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (356mg, 0.938mmol, 49.5% yield), it is pale solid, LCMS (m/z):374.3[M+H ]⁺。

¹H NMR(400MHz,CDCl₃):δ 13.81 (s, 1H), 9.57 (d, J=1.4Hz, 1H), 8.99 (dd, J=2.1, 0.8Hz, 1H), 8.52 (dd, J=2.0,0.9Hz, 1H), 8.47 (m, J=2.2,1.1Hz, 1H), 8.32-8.28 (m, 2H), 7.64 (d, J=8.0Hz, 1H), 7.47 (d, J=8.0Hz, 1H), 5.71 (dd, J=6.1,3.2Hz, 1H), 3.35-3.16 (m, 3H), 3.03 (dd, J=12.1,3.3Hz, 1H), 2.51 (s, 3H), 2.35 (dddd, J=14.0,9.9,6.1,4.0Hz, 1H), 2.15-2.03(m,1H)。

Embodiment 155

Synthesize (4S) -7- (1- methyl isophthalic acid H- pyrazoles -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To with the chloro- N- of argon-degassed 20min (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (600mg, 1.894mmol), (1- methyl isophthalic acid H- pyrazoles -4- bases) boric acid (286mg, 2.273mmol) and K₃PO₄(804mg, 3.79mmol) is molten in 1,4- dioxanes (12.00mL), water (4.00mL) X-Phos (90mg, 0.189mmol) and three (dibenzalacetone) two palladium (0) (87mg, 0.095mmol) is added in liquid, again Use argon-degassed 10min.Reactant mixture is stirred into 4h at 100 DEG C, room temperature is subsequently cooled to.By reactant mixture through diatomite Filter, then filtrate water dilutes and extracted (3 × 10mL) with EtOAc.Organic layer water, the aqueous salt solu-tion of merging, warp Sodium sulphate is dried and evaporated, and obtains crude compound (TLC eluant, eluents：5%MeOH/DCM:R_f-0.3；UV activation).Roughization Compound is purified through column chromatography, is eluted using 100-200 silica gel mesh and with 1.5-3%MeOH/DCM, obtains pure (4S) -7- (1- Methyl isophthalic acid H- pyrazoles -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (354mg, 0.963mmol, 50.9% yield), it is faint yellow solid, LCMS (m/z):363.1 [M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ 14.09 (s, 1H), 9.39 (d, J=1.5Hz, 1H), 8.51 (d, J= 0.8Hz, 1H), 8.47 (dd, J=2.6,1.6Hz, 1H), 8.37-8.36 (m, 1H), 8.26 (d, J=0.8Hz, 1H), 7.56 (d, J=8.0Hz, 1H), 7.34 (d, J=8.0Hz, 1H), 5.46 (dd, J=6.0,3.1Hz, 1H), 3.92 (s, 3H), 3.22- 3.13 (m, 1H), 3.12-3.02 (m, 2H), 2.94 (dd, J=12.0,3.3Hz, 1H), 2.24 (dddd, J=13.6,9.8, 6.1,3.8Hz, 1H), 1.92 (m, J=14.4,7.4Hz, 1H).

Embodiment 156

Synthesize (4S) -7- (6- ethylpyridine -3- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines with argon-degassed 20min And [2,3-b] [1,4] diaza- 5 (2H)-formamides (600mg, 1.894mmol), (6- ethylpyridine -3- bases) boric acid (343mg, 2.273mmol) and K₃PO₄(804mg, 3.79mmol) is molten in 1,4- dioxanes (12.00mL), water (3.00mL) X-Phos (90mg, 0.189mmol) and three (dibenzalacetone) two palladium (0) (87mg, 0.095mmol) is added in liquid, again Use argon-degassed 10min.Reactant mixture is stirred into 4h at 100 DEG C, room temperature is subsequently cooled to.By reactant mixture through diatomite Filter, then filtrate water dilutes and extracted (3 × 10mL) with EtOAc.Organic layer water, the aqueous salt solu-tion of merging, warp Sodium sulphate is dried and evaporated, and obtains crude compound (TLC eluant, eluents：5%MeOH/DCM:R_f-0.3；UV activation).Roughization Compound is purified through column chromatography, is eluted using 100-200 silica gel mesh and with 1.5-3%MeOH/DCM, obtains pure (4S) -7- (6- Ethylpyridine -3- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (354mg, 0.883mmol, 46.6% yield), it is pale solid, LCMS (m/z):388.3[M+H ]⁺。

¹H NMR(400MHz,CDCl₃):δ 13.77 (s, 1H), 9.54 (d, J=1.5Hz, 1H), 9.22-9.01 (d, 1H), 8.47 (dd, J=2.5Hz, 1H), 8.35 (d, J=2.4Hz, 1H), 8.29 (d, J=2.6Hz, 1H), 7.63 (d, J=8.0Hz, 1H), 7.42 (d, J=8.0Hz, 1H), 7.35-7.32 (d, 1H), 5.70 (dd, J=6.0,3.2Hz, 1H), 3.39-3.13 (m, 3H), 3.02 (dd, J=12.1,3.3Hz, 1H), 2.92 (q, J=7.6Hz, 2H), 2.35 (dddd, J=13.9,9.9,6.1, 4.0Hz, 1H), 2.21-1.98 (m, 1H), 1.38 (t, J=7.6Hz, 3H).

Embodiment 157

Synthesize (4S)-N- (1H- indazole -5- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaIn the solution of (600mg, 2.378mmol) in THF (30ml) add triphosgene (353mg, 1.189mmol), then stir to room temperature, continue 1h.Then in succession room temperature add 1H- indazole -5- amine (412mg, 3.09mmol) with triethylamine (1.657mL, 11.89mmol).Reactant mixture is heated into 16h at 70 DEG C in seal pipe, poured into Saturation NaHCO₃Solution (180mL), is extracted with ethyl acetate (3x50mL).The organic layer of merging is through anhydrous Na₂SO₄Dry and subtract Pressure concentration, obtains crude product.Crude mixture purifies (ammonium hydrogen carbonate and acetonitrile) through preparation HPLC, obtains (4S)-N- (1H- Indazole -5- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (210mg, 0.489mmol, 20.54% yield), it is Light brown solid (TLC:5%MeOH/DCM, R_f=0.3), LCMS (m/z):412.27[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 12.98 (s, 1H), 12.92 (s, 1H), 8.65 (d, J=5.04Hz, 1H), 8.09-7.94 (d, J=4.5Hz, 1H), 7.58-7.47 (m, 2H), 7.47-7.41 (m, 2H), 7.41-7.32 (d, J= 8.4Hz, 1H), 7.32-7.17 (d, J=8.3Hz, 1H), 5.74 (dd, J=5.70,3.07Hz, 1H), 3.37-3.11 (m, 3H), 3.11-2.86 (m, 1H), 2.60 (s, 3H), 2.22 (dt, J=14.03,7.02Hz, 2H), 1.99-1.81 (m, 1H).

Embodiment 158

Synthesize (4S) -7- (2- picoline -4- bases)-N- (pyridazine -3- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 30 DEG C to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaIn the solution of the stirring of (700mg, 2.77mmol) in THF (15mL) add triphosgene (412mg, 1.387mmol) and stirring 1h.Then 30 DEG C add pyridazine -3- amine (792mg, 8.32mmol) and triethylamine (1.160mL, 8.32mmol), reactant mixture is heated into 16h at 75 DEG C.So that 30 DEG C of reactant mixture and pouring into cold water (30mL), use DCM extracts (2x35ml).The organic layer of merging salt water washing, it is dried over sodium sulfate and solvent is evaporated under reduced pressure, obtain crude product. Crude mixture is through flash column chromatography (silica gel：100-200 mesh, solvent system) and preparation HPLC ((10mM ammonium hydrogen carbonate (water Solution) MP-B：Acetonitrile post：μ methods-the t%b 0/60,13.5/60,14/100,17/ of xbridge C18 (150x30) mm 5 100,17.1/60 flow velocity：30ml/min, eluant, eluent：THF+ACN.+ water) purifying, obtain (4S) -7- (2- picolines -4- Base)-N- (pyridazine -3- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine (270mg, 0.723mmol, 25.7%), it is faint yellow solid (TLC:R_f:0.4,10%MeOH/EtOAc), LCMS (m/ z):374.26[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 14.16 (s, 1H), 8.94 (dd, J=4.7,1.5Hz, 1H), 8.66 (d, J= 5.3Hz, 1H), 8.48 (dd, J=9.1,1.5Hz, 1H), 8.38-830 (m, 1H), 8.22 (s, 1H), 7.83-7.50 (m, 3H), 5.69 (dd, J=6.0,3.2Hz, 1H), 3.41-3.16 (m, 3H), 3.03 (dd, J=12.1,3.3Hz, 1H), 2.81 (s, 3H), 2.35 (dddd, J=13.9,10.0,6.1,4.0Hz, 1H), 2.22-2.00 (m, 1H).

Embodiment 159

Synthesize (4S) -7- (2,5- lutidines -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

28 DEG C by tripotassium phosphate (804mg, 3.79mmol) add to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (600mg, 1.894mmol), 2,5- diformazans Base -4- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) pyridine (662mg, 2.84mmol) is in 1,4- bis- Evil In alkane (10mL) and the solution of the stirring in water (1.667mL).Reactant mixture is deaerated 15min, Pd is added₂(dba)₃ (87mg, 0.095mmol) and X-phos (90mg, 0.189mmol).Reactant mixture is deaerated 15min again, and will reaction mixing Thing stirs 3hr at 100 DEG C.Reactant mixture is cooled to 28 DEG C, then distributed between water (10mL) and EtOAc (25mL). Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain thick material, be that (TLC is eluted pink solid System:10% methanol/EtOAc；R_f=0.3).Thick material is purified through column chromatography, uses (100-200 mesh) silica gel, and use 5-8% Methanol/EtOAc elutions, obtains pure (4S) -7- (2,5- lutidines -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(337mg, 0.860mmol, 45.4% are received -5 (2H)-formamides Rate), it is pale solid, LCMS (m/z):388.19[M+H]⁺。

¹H NMR(CDCl₃,400MHz):δ 13.48 (s, 1H), 9.49 (d, J=1.6Hz, 1H), 8.46 (s, 1H), 8.34- 8.10 (m, 2H), 7.63 (d, J=7.8Hz, 1H), 7.45 (s, 1H), 7.15 (d, J=7.9Hz, 1H), 5.71 (dd, J=6.0, 3.2Hz, 1H), 3.51-3.09 (m, 3H), 3.04 (dd, J=12.1,3.3Hz, 1H), 2.63 (s, 3H), 2.43 (s, 3H), 2.40-2.27(m,1H),2.20-1.96(m,1H)。

Embodiment 160

Synthesis (4S) -7- morpholinoes-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides

To the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (600mg, 1.894mmol) and morpholine (0.330mL, 3.79mmol) are in 1,4- dioxanes (6mL) Cs is added in the solution of degassing₂CO₃(1234mg, 3.79mmol), x-phos (361mg, 0.758mmol) and Pd (OAc)₂ (85mg,0.379mmol).Reactant mixture is stirred into 16h at 110 DEG C.Reactant mixture is poured into cold water (50mL) and second is used Acetoacetic ester extracts (2x50mL).The organic layer of merging is concentrated under reduced pressure through anhydrous sodium sulfate drying, obtains crude product.Crude product is passed through Flash column chromatography (silica gel：100-200 mesh, eluant, eluent：2%MeOH/DCM), obtain (4S) -7- morpholinoes-N- (pyrazine - 2- yls) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (268mg, 0.707mmol, 37.3% yield), it is faint yellow solid (TLC:R_f:0.2, net EtOAc), LCMS (m/z):368[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 13.41 (s, 1H), 9.53 (d, J=1.5Hz, 1H), 8.37-7.90 (m, 2H), 7.39 (d, J=8.6Hz, 1H), 6.29 (d, J=8.6Hz, 1H), 5.62 (dd, J=6.0,3.2Hz, 1H), 3.98-3.72 (m, 4H), 3.62-3.46 (m, 4H), 3.22 (dddd, J=12.2,8.5,3.7,2.2Hz, 1H), 3.18-3.05 (m, 2H), 2.93 (dd, J=11.8,3.3Hz, 1H), 2.27 (dd, J=10.1,4.0Hz, 1H), 2.01 (d, J=6.9Hz, 1H).

Embodiment 161

Synthesize (4S)-N- (5- ethyl pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature by (5- ethyl pyrazine -2- bases) phenyl carbamates (1446mg, 5.94mmol) and DMAP (726mg, 5.94mmol) add to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diazaThe solution of the stirring of (500mg, 1.982mmol) in tetrahydrofuran (THF) (15mL), then will reaction Mixture stirs 16h at 80 DEG C.Reactant mixture is reached room temperature, (2X30mL) is diluted with water (40mL) and salt with ethyl acetate Water (20mL) is washed.Organic layer is separated, it is then dried over sodium sulfate, filter and concentrate.Residue is purified through column chromatography, uses silicon Glue (100-200 mesh), 1% methanol/DCM obtains required product as eluant, eluent.Compound is tied again using ethanol and pentane Crystalline substance, obtains (4S)-N- (5- ethyl pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro-Isosorbide-5-Nitrae-endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides (245mg, 0.602mmol, 30.4% yield), it is solid for canescence Body.(TLC:5% methanol/DCM.R_f:0.4UV), LCMS (m/z):402.3[M+H]⁺。

¹H NMR(400MHz,CDCl₃) δ 13.46 (s, 1H), 9.37 (s, 1H), 8.67 (dd, J=5.3,0.8Hz, 1H), 8.20 (d, J=0.6Hz, 1H), 7.98 (ddd, J=5.4,1.8,0.7Hz, 1H), 7.74-7.54 (m, 2H), 7.48 (d, J= 7.9Hz, 1H), 5.72 (dd, J=6.0,3.2Hz, 1H), 3.37-3.17 (m, 1H), 3.03 (dd, J=12.1,3.3Hz, 1H), 2.80 (q, J=7.6Hz, 2H), 2.62 (s, 3H), 2.35 (dddd, J=14.0,9.9,6.1,4.1Hz, 1H), 2.22-1.92 (m, 3H), 1.32 (t, J=7.6Hz, 3H).

Embodiment 162

Synthesize (4S) -7- (2- Jia Ji oxazole -5- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides:

28 DEG C by tripotassium phosphate (804mg, 3.79mmol) add to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (600mg, 1.894mmol), 2- methyl- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- base) oxazoles (475mg, 2.273mmol) are in 1,4- dioxanes by 5- In (10mL) and the solution of the stirring in water (1.667mL).Reactant mixture is deaerated 15min, Pd is added₂(dba)₃(87mg, 0.095mmol) with X-phos (90mg, 0.189mmol).Reactant mixture is deaerated 15min again, and reactant mixture is existed 100 DEG C of stirring 3hr.Reactant mixture is cooled to 28 DEG C, then distributed between water (10mL) and EtOAc (25mL).Separation Organic layer, then through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain thick material, it is that (TLC is eluted brown solid Agent：10%MeOH/EtOAc；R_f=0.2；UV activation).Thick material is purified by preparation HPLC, desired product is obtained (4S) -7- (2- Jia Ji oxazole -5- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (153mg, 0.418mmol, 22.04% yield), it is white solid, LCMS (m/z): 364.32[M+H]⁺。

¹H NMR(CDCl₃,400MHz):δ 13.72 (s, 1H), 9.54 (d, J=1.5Hz, 1H), 8.44-8.17 (m, 2H), 8.07 (s, 1H), 7.57 (d, J=7.9Hz, 1H), 7.30-7.13 (m, 1H), 5.67 (dd, J=6.0,3.2Hz, 1H), 3.45- 3.08 (m, 3H), 3.01 (dd, J=12.1,3.3Hz, 1H), 2.58 (s, 3H), 2.34 (dddd, J=14.1,9.9,6.1, 4.2Hz,1H),2.22-1.95(m,1H)。

Embodiment 163

Synthesize (4S) -7- (2- picoline -4- bases)-N- (pyrimidine -5- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 30 DEG C to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaIn the solution of the stirring of (700mg, 2.77mmol) in THF (15mL) add triphosgene (823mg, 2.77mmol) and stirring 1h.Then pyrimidine -5- amine (317mg, 3.33mmol) and TEA (0.387mL, 2.77mmol) are added, will Reactant mixture heats 16h at 75 DEG C.So that reactant mixture reaches 30 DEG C and poured into cold water (30mL), extracted with DCM (2x50ml).The organic layer of merging salt water washing, it is dried over sodium sulfate and solvent is evaporated under reduced pressure, obtain crude product.It is rough mixed Compound is through flash column chromatography (silica gel：100-200 mesh, 3%MeOH/ ethyl acetate), obtain (4S) -7- (2- picolines - 4- yls)-N- (pyrimidine -5- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first Acid amides (275mg, 0.737mmol, 32%), it is faint yellow solid (TLC:R_f:0.4,10%MeOH/EtOAc), LCMS (m/ z):374.22[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 13.25 (s, 1H), 9.01 (s, 2H), 8.94 (s, 1H), 8.68 (d, J= 0.8Hz, 1H), 7.67 (d, J=7.9Hz, 1H), 7.55 (dt, J=1.6,0.8Hz, 1H), 7.47 (ddd, J=5.2,1.7, 0.7Hz, 1H), 7.39 (s, 1H), 5.69 (dd, J=5.9,3.2Hz, 1H), 3.33-3.13 (m, 3H), 3.04 (dd, J= 12.2,3.2Hz, 1H), 2.69 (s, 3H), 2.36 (dd, J=10.1,4.1Hz, 1H), 2.15-2.04 (m, 1H).

Embodiment 164

Synthesize (4S)-N- (5- cyclopropyl pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature by (5- cyclopropyl pyrazine -2- bases) phenyl carbamates (1214mg, 4.76mmol) and DMAP (581mg, 4.76mmol) adds to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diazaMixed in the solution of the stirring of (400mg, 1.585mmol) in THF (15mL) and by reaction Thing stirs 16h at 80 DEG C.Reactant mixture is reached room temperature, (2X30mL) is diluted with ethyl acetate, with water (20mL) and salt solution (20mL) is washed.Organic layer is separated, it is then dried over sodium sulfate, filter and concentrate.Residue is purified through column chromatography, uses silica gel (100-200 mesh), 2% methanol/DCM obtains required product as eluant, eluent.Compound is recrystallized by ethanol and pentane, Obtain (4S)-N- (5- cyclopropyl pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides (260mg, 0.623mmol, 45.2%), it is pale solid, (R_f:0.4,TLC:5% methanol/DCM), LCMS (m/z):414.3[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 13.59 (s, 1H), 9.35 (d, J=1.5Hz, 1H), 8.62 (dd, J=5.3, 0.8Hz, 1H), 8.20 (d, J=1.5Hz, 1H), 8.07 (q, J=0.9Hz, 1H), 7.71-7.59 (m, 2H), 7.48 (d, J= 8.0Hz, 1H), 5.71 (dd, J=6.0,3.2Hz, 1H), 3.37-3.10 (m, 3H), 3.02 (dd, J=12.1,3.3Hz, 1H), 2.74 (s, 3H), 2.34 (dd, J=10.1,4.1Hz, 1H), 2.07 (d, J=8.0Hz, 1H), 1.21-0.66 (m, 5H).

Embodiment 165

Synthesize (4S) -7- (2- methylpyrimidine -5- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

Tripotassium phosphate (804mg, 3.79mmol) is added into the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (600mg, 1.894mmol), (2- methylpyrimidines -5- Base) stirring of the boric acid (314mg, 2.273mmol) in 1,4- dioxanes (12mL) and water (2.000mL) solution in.Will be anti- Mixture degassing 15min is answered, X-phos (90mg, 0.189mmol) and Pd is added₂(dba)₃(87mg,0.095mmol).Will reaction Mixture deaerates 15min again, then stirs 6hr at 100 DEG C.Reactant mixture is cooled to 28 DEG C, then distributed in water Between (30mL) and EtOAc (70mL).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filtering.Evaporation filtrate obtains thick thing Matter (TLC eluant, eluents：5%MeOH/CHCl₃:R_f=0.2；UV activation).Crude compound passes through through 100-200 silica gel purifications 20%MeOH/ ethyl acetate is eluted, and obtains (4S) -7- (2- methylpyrimidine -5- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(353mg, 0.938mmol, 49.5% are received -5 (2H)-formamides Rate), it is faint yellow solid, LCMS (m/z):375.19[M+H]⁺。

¹H NMR(CDCl₃,400MHz):δ 13.57 (s, 1H), 9.53 (d, J=1.5Hz, 1H), 9.33 (s, 2H), 8.38 (d, J=1.6Hz, 1H), 8.30 (s, 1H), 7.66 (d, J=8.0Hz, 1H), 7.40 (d, J=8.0Hz, 1H), 5.71 (dd, J =5.9,3.2Hz, 1H), 3.41-3.11 (m, 3H), 3.04 (dd, J=12.1,3.3Hz, 1H), 2.84 (s, 3H), 2.36 (dddd, J=14.0,9.9,6.1,4.1Hz, 1H), 2.11 (d, J=7.1Hz, 1H).

Embodiment 166

Synthesize (4S)-N- (6- cyclopropyl pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature to 7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diazaIn the solution of the stirring of (600mg, 0.198mmol) in THF (30mL) add triphosgene (353mg, 0.099mmol) and stirring 30min.Then 6- cyclopropyl pyrazine -2- amine (321mg, 2.378mmol) and Et are added in room temperature₃N (1.657mL,0.991mmol).Reactant mixture is stirred into 16h at 65 DEG C.Reactant mixture is poured into NaHCO₃In solution simultaneously Use CH₂Cl₂Extract (3x50mL).The organic layer of merging is through anhydrous Na₂SO₄Dry and be concentrated under reduced pressure, obtain crude product.It is rough mixed Compound is through flash column chromatography (silica gel：100-200 mesh, uses 2%MeOH/CH₂Cl₂Elution), obtain (4S)-N- (6- cyclopropyl Pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (208mg, 0.479mmol, 30.2% yield), it is white solid (TLC:10%MeOH/CH₂Cl₂, R_f:0.5), LCMS (m/z):414.29[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.11 (s, 1H), 9.29 (s, 1H), 8.80 (d, J=5.26Hz, 1H), 8.57 (d, J=4.60Hz, 1H), 8.15 (s, 1H), 7.97-7.67 (m, 1H), 7.64 (d, J=7.89Hz, 1H), 7.59- 7.42 (m, 1H), 5.72 (dd, J=5.92,3.29Hz, 1H), 3.34-3.10 (m, 3H), 3.39-3.09 (m, 1H), 2.62 (s, 3H),2.44-2.18(m,1H),2.17–1.97(m,1H),1.13–0.93(m,4H)。

Embodiment 167

(4S) -7- (2- picoline -4- bases)-N- (1H-1,2,3- triazole-4-yls) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Triphosgene (0.588g, 1.982mmol) is added into (4S) -7- (2- picoline -4- bases) -2,3,4,5- at 0 DEG C Tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(0.5g, 1.982mmol) is in tetrahydrofuran (50mL) Stirring solution in, triethylamine (0.829mL, 5.94mmol) and 1H-1,2,3- triazole -4- amine salt acid are then added at 0 DEG C Salt (0.597g, 4.95mmol).Reactant mixture is stirred into 48h at 70 DEG C.Reactant mixture is cooled to 28 DEG C, then distributed Between water (50mL) and EtOAc (50mL).Separate organic layer and with water and salt water washing.Organic layer is through anhydrous Na₂SO₄Dry, Filter and evaporate filtrate, obtain rough yellow solid (TLC eluant, eluents：10%MeOH/EtOAc:R_f=0.35；UV is activated ).Crude compound is eluted with 5%MeOH/ ethyl acetate through 100-200 silica gel purifications, obtains (4S) -7- (2- methyl pyrroles Pyridine -4- bases)-N- (1H-1,2,3- triazole-4-yls) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (0.21g, 0.558mmol, 28.2% yield), it is light yellow solid, LCMS (m/z):363.24[M +H]⁺。

¹H NMR(400MHz,DMSO-d6):δ 14.58 (s, 1H), 13.55 (s, 1H), 8.58 (d, J=5.2Hz, 1H), 7.94 (s, 2H), 7.84-7.75 (m, 2H), 7.71 (d, J=8.0Hz, 1H), 5.47 (dd, J=5.8,3.0Hz, 1H), 3.27- 3.16 (m, 1H), 3.16-3.01 (m, 2H), 2.97 (dd, J=12.0,3.2Hz, 1H), 2.64 (s, 3H), 2.24 (dddd, J= 13.7,9.8,5.9,3.7Hz, 1H), 1.94 (dddd, J=14.5,8.7,4.5Hz, 1H).

Embodiment 168

Synthesize (4S) -7- (2- picoline -4- bases)-N- ((S) -1- (pyridine -2- bases) ethyl) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diazaIn the solution of (500mg, 1.982mmol) in tetrahydrofuran (10mL) add triphosgene (588mg, 1.982mmol) with triethylamine (1.657mL, 11.89mmol).30min is stirred at room temperature in reactant mixture, (S) -1- is added (pyridine -2- bases) ethamine, then stirs 16h at 70 DEG C.Removal of solvent under reduced pressure and be diluted with water and be extracted with ethyl acetate (2 × 50mL).The organic layer of merging is washed with water (30mL), saturated brine solution.Organic layer is through anhydrous sodium sulfate drying, and filtering simultaneously will Filtrate is evaporated, and is obtained crude product, is brown solid.(TLC eluant, eluents：10% ethanol/methylene, R_f=0.3；UV is activated ).Crude compound is purified (neutral alumina) through column chromatography, and product is eluted with 55-60% ethyl acetate/hexanes.It will collect Fraction be evaporated under reduced pressure, obtain pure (4S) -7- (2- picoline -4- bases)-N- ((S) -1- (pyridine -2- bases) ethyl) -3, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (277mg, 0.684mmol, 34.5% yield), it is pale solid, LCMS (m/z):401.2[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 11.09 (d, J=7.6Hz, 1H), 8.56 (dd, J=5.3,0.8Hz, 1H), 8.36 (ddd, J=4.8,1.8,0.9Hz, 1H), 7.78 (dt, J=1.4,0.7Hz, 1H), 7.63-7.60 (m, 2H), 7.59 (dd, J=7.7,1.9Hz, 1H), 7.31-7.29 (m, 2H), 7.11 (ddd, J=7.6,4.8,1.2Hz, 1H), 5.64 (dd, J =6.0,3.2Hz, 1H), 5.32-5.23 (m, 1H), 3.27-3.06 (m, 3H), 2.95 (dd, J=12.0,3.3Hz, 1H), 2.60 (s, 3H), 2.22 (dd, J=10.0,4.0Hz, 1H), 2.03-1.89 (m, 1H), 1.62 (d, J=6.9Hz, 3H).

Embodiment 169

Synthesize (4S) -7- (2- picoline -4- bases)-N- ((R) -1- (pyridine -2- bases) ethyl) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaIn the solution of (500mg, 1.982mmol) in tetrahydrofuran (10mL) add triphosgene (588mg, 1.982mmol) with triethylamine (1.657mL, 11.89mmol).30min is stirred at room temperature in reactant mixture, (R) -1- is added (pyridine -2- bases) ethamine (726mg, 5.94mmol) and 70 DEG C stir 16h.Removal of solvent under reduced pressure, is diluted with water (15mL), and It is extracted with ethyl acetate (2 × 50mL).The organic layer of merging is washed with water (30mL), saturated brine solution, then through anhydrous sulphur Sour sodium is dried.Solution is filtered and concentrated, crude product is obtained, is brown solid.(TLC eluant, eluents：10% ethanol/methylene, R_f=0.3；UV activation).Crude compound through column chromatography purify (neutral alumina) product 55-60% ethyl acetate/oneself Alkane is eluted, and obtains (4S) -7- (2- picoline -4- bases)-N- ((R) -1- (pyridine -2- bases) ethyl) -3,4- dihydros-Isosorbide-5-Nitrae-bridge Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (240mg, 0.595mmol, 30.0% yield), its For yellow jelly, LCMS (m/z):401.2[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 11.06 (d, J=7.5Hz, 1H), 8.57 (dd, J=5.2,0.8Hz, 1H), 8.43 (ddd, J=4.8,1.8,0.9Hz, 1H), 7.78 (q, J=0.9Hz, 1H), 7.63 (ddd, J=5.1,1.9,0.7Hz, 1H), 7.61 (dd, J=7.8,1.9Hz, 1H), 7.55 (d, J=7.9Hz, 1H), 7.36-7.30 (m, 2H), 7.13 (ddd, J= 7.6,4.8,1.2Hz, 1H), 5.62 (dd, J=5.9,3.2Hz, 1H), 5.27 (p, J=7.0Hz, 1H), 3.30-3.21 (m, 1H), 3.18 (ddd, J=12.2,9.9,6.9Hz, 1H), 3.06 (dt, J=12.0,2.1Hz, 1H), 2.92 (dd, J=11.9, 3.3Hz, 1H), 2.60 (s, 3H), 2.27 (dddd, J=13.9,9.9,6.1,3.9Hz, 1H), 2.07 (dt, J=14.0, 7.5Hz, 1H), 1.62 (d, J=6.9Hz, 3H).

Embodiment 170

Synthesis (4S)-N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (3- (trifluoromethyl) phenyl) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

0 DEG C to (4S)-N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine - 4- yls) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 4.0M hydrochloric acid dioxane solutions (10mL, 1.080mmol) are added in the solution of (2H)-formamide (600mg, 1.080mmol). 3h is stirred at room temperature in reactant mixture.The solvent of reactant mixture is evaporated, is then neutralized and is extracted with EtOAc with sodium acid carbonate (3 × 50mL), the organic layer aqueous salt solu-tion of merging is then dried over sodium sulfate, and evaporated, obtains the rough chemical combination of 350mg Thing.Crude product adds to silicagel column and eluted with 2%MeOH/DCM, and the fraction of collection is evaporated, obtain (4S)-N- (2- ((S) -2, 3- dihydroxy propoxyl group) pyridin-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza- 5 (2H)-formamides (253mg, 0.487mmol, 45.1% yield), it is pale solid. (TLC:R_f:0.2,5%MeOH/DCM), LCMS (m/z):516.3[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆)δppm 13.02(s,1H),8.08-8.33(m,2H),7.77-8.00(m, 3H), 7.76-7.59 (m, 2H), 7.11 (d, J=1.53Hz, 1H), 6.80 (dd, J=5.70,1.75Hz, 1H), 5.47 (dd, J =5.92,3.07Hz, 1H), 4.85 (d, J=5.26Hz, 1H), 4.58 (t, J=5.70Hz, 2H), 4.23 (dd, J=10.85, 4.49Hz, 1H), 4.12 (dd, J=10.74,6.14Hz, 1H), 3.78 (dq, J=10.69,5.43Hz, 1H), 3.53-3.32 (m, 1H), 3.25-3.05 (m, 3H), 2.97 (dd, J=11.95,3.18Hz, 2H), 2.37-2.18 (m, 1H), 2.03-1.86 (m,1H)。

Embodiment 171

Synthesize (4S) -7- (5,6- lutidines -3- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

28 DEG C by tripotassium phosphate (804mg, 3.79mmol) add to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (600mg, 1.894mmol), (5,6- bis- Picoline -3- bases) stirring of the boric acid (343mg, 2.273mmol) in 1,4- dioxanes (10mL) and water (1.667mL) In solution.Reactant mixture is deaerated 15min, Pd is added₂(dba)₃(87mg, 0.095mmol) and X-phos (90mg, 0.189mmol).Reactant mixture is deaerated 15min again, and reactant mixture is stirred into 3hr at 100 DEG C.By reactant mixture 28 DEG C are cooled to, is then distributed between water (10mL) and EtOAc (25mL).Organic layer is separated, then through anhydrous Na₂SO₄It is dry It is dry, filter and evaporate filtrate, obtain thick material, it is brown solid (TLC eluant, eluents：10%MeOH/EtOAc；R_f=0.3； UV activation).Thick material is purified through column chromatography, is used (100-200 mesh) silica gel and is eluted with 5-10%MeOH/EtOAc, is obtained Pure (4S) -7- (5,6- lutidines -3- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza- 5 (2H)-formamides (420mg, 1.078mmol, 56.9% yield), it is pale solid, LCMS

(m/z):388.19[M+H]⁺。

¹H NMR(CDCl₃,400MHz):δ 13.85 (s, 1H), 9.57 (d, J=1.4Hz, 1H), 8.86 (d, J=2.3Hz, 1H), 8.54-8.34 (m, 1H), 8.34-8.19 (m, 2H), 7.62 (d, J=8.0Hz, 1H), 7.44 (d, J=8.0Hz, 1H), 5.71 (dd, J=6.0,3.2Hz, 1H), 3.45-3.08 (m, 3H), 3.02 (dd, J=12.1,3.3Hz, 1H), 2.59 (s, 3H), 2.47 (d, J=0.8Hz, 3H), 2.35 (dddd, J=14.0,10.0,6.1,4.0Hz, 1H), 2.22-2.00 (m, 1H).

Embodiment 172

Synthesize (4S) -7- (3- (dimethylamino) phenyl)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides:

28 DEG C by tripotassium phosphate (804mg, 3.79mmol) add to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (600mg, 1.894mmol), (3- (diformazans Base amino) phenyl) stirring of the boric acid (469mg, 2.84mmol) in 1,4- dioxanes (10mL) and water (1.667mL) solution In.Reactant mixture is deaerated 15min, Pd is added₂(dba)₃(87mg, 0.095mmol) and X-phos (90mg, 0.189mmol).Reactant mixture is deaerated 15min again, and reactant mixture is stirred into 3hr at 100 DEG C.By reactant mixture 28 DEG C are cooled to, is then distributed between water (10mL) and EtOAc (25mL).Organic layer is separated, then through anhydrous Na₂SO₄It is dry It is dry, filter and evaporate filtrate, obtain thick material, it is brown solid (TLC eluant, eluents：10%MeOH/EtOAc；R_f=0.5；UV Activation).Thick material is purified by using column chromatography, (100-200 mesh) silica gel is used and is washed with 0-5%MeOH/EtOAc It is de-, obtain pure (4S) -7- (3- (dimethylamino) phenyl)-N- (pyrazine -2- bases) -3,4- dihydro-Isosorbide-5-Nitrae-endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (273mg, 0.660mmol, 34.8% yield), it is canescence Solid, LCMS (m/z):402.26[M+H]⁺。

¹H NMR(DMSO-d₆,400MHz):δ 13.66 (s, 1H), 9.39 (s, 1H), 8.35 (q, J=2.7Hz, 2H), 7.81-7.42 (m, 1H), 7.44-7.19 (m, 4H), 6.92-6.77 (m, 1H), 5.53 (dd, J=5.9,3.1Hz, 1H), 3.29-3.00 (m, 3H), 2.98 (s, 7H), 2.25 (ddd, J=13.8,10.3,5.4Hz, 1H), 1.96 (dt, J=14.9, 7.8Hz,1H)。

Embodiment 173

Synthesize ((4S) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] Diaza- 5 (2H)-yls) (pyrrolidin-1-yl) ketone

It is sub- to (4S) -7- (2- picoline -4- the bases) -2,3,4,5- tetrahydrochysene -1,4- bridges being stirred at room temperature under a nitrogen Picoline simultaneously [2,3-b] [1,4] diazaThe solution of (500mg, 1.982mmol) in tetrahydrofuran (THF) (10mL) Middle addition triphosgene (294mg, 0.991mmol), is then stirred at room temperature 30min.After 30 minutes, triethylamine is added (1.381mL, 9.91mmol) and pyrrolidines (183mg, 2.58mmol).Reactant mixture is stirred into 16hr at 60 DEG C.Reaction is mixed Compound is gone out with 2x15ml water quenchings and uses 2x20ml ethyl acetate to extract, and organic layer is washed with water, then dried over sodium sulfate and subtract Pressure concentration, obtains crude compound.Crude product is eluted through 100-200 silica gel purifications post and with 4% methanol/DCM, obtains pure change Compound ((4S) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-yls) (pyrrolidin-1-yl) ketone (350mg, 0.993mmol, 50.1% yield), it is yellow solid (R_fValue： 0.4,10% methanol/DCM), LCMS (m/z):350.25[M+H]⁺。

¹H NMR (400MHz, DMSO-d6) δ 8.49 (dd, J=5.2,0.8Hz, 1H), 7.75 (dt, J=1.4,0.7Hz, 1H), 7.67 (ddd, J=5.4,1.7,0.7Hz, 1H), 7.50 (d, J=7.9Hz, 1H), 7.44 (d, J=7.9Hz, 1H), 4.42 (d, J=3.8Hz, 1H), 3.44-3.24 (m, 4H), 3.14 (ddt, J=12.0,6.1,3.3Hz, 1H), 3.03 (dd, J =13.8,5.2Hz, 2H), 2.89 (dd, J=11.6,3.3Hz, 1H), 2.55 (s, 3H), 2.23-2.01 (m, 2H), 1.99- 1.86(m,4H).

Embodiment 174

Synthesize (4- methylpiperazine-1-yls) ((4S) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-yls) ketone

It is sub- to (4S) -7- (2- picoline -4- the bases) -2,3,4,5- tetrahydrochysene -1,4- bridges stirred under a nitrogen in room temperature Picoline simultaneously [2,3-b] [1,4] diazaThe solution of (500mg, 1.982mmol) in tetrahydrofuran (THF) (10mL) Middle addition triphosgene (294mg, 0.991mmol), is then stirred at room temperature 30min.After 30 minutes, triethylamine is added (1.381mL, 9.91mmol) and 1- methyl piperazines (258mg, 2.58mmol).Reactant mixture is stirred into 16hr at 60 DEG C.Make Obtain reactant mixture to reach room temperature and gone out with 2x15ml water quenchings and use 2x20ml ethyl acetate to extract, organic layer is washed with water, so It is dried over sodium sulfate afterwards and be concentrated under reduced pressure, obtain crude compound 500mg.Purified on column chromatography is purified, and obtains pure chemical combination Thing (4- methylpiperazine-1-yls) ((4S) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza- 5 (2H)-yls) ketone (280mg, 0.725mmol, 36.6% yield), it is yellow solid, (R_fValue： 0.25,10% methanol/DCM), LCMS (m/z):379.1[M+H]⁺。

¹H NMR (400MHz, DMSO-d6) δ 8.51 (dd, J=5.3,0.8Hz, 1H), 7.83-7.72 (m, 1H), 7.70 (ddd, J=5.2,1.7,0.7Hz, 1H), 7.53-7.36 (m, 2H), 4.36 (d, J=3.9Hz, 1H), 3.60 (s, 1H), 3.42 (s, 2H), 3.22-2.90 (m, 3H), 2.61-2.44 (m, 9H), 2.15 (d, J=10.0Hz, 5H).

Embodiment 175

Synthesis (4S)-N- cyclopenta -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza- 5 (2H)-formamides

It is sub- to (4S) -7- (2- picoline -4- the bases) -2,3,4,5- tetrahydrochysene -1,4- bridges stirred under a nitrogen in room temperature Picoline simultaneously [2,3-b] [1,4] diazaThe solution of (500mg, 1.982mmol) in tetrahydrofuran (THF) (10mL) Middle addition triphosgene (294mg, 0.991mmol), is then stirred at room temperature 30min.Thereto add triethylamine (1.381mL, 9.91mmol) with cyclopenta amine (219mg, 2.58mmol).Reactant mixture is stirred into 16hr at 60 DEG C.Reactant mixture is used 2x15ml water quenchings are gone out and use 2x20ml ethyl acetate to extract, and organic layer is washed with water, then dried over sodium sulfate and be concentrated under reduced pressure, Obtain crude compound 500mg.Purified on column chromatography is purified, and obtains pure compound (4S)-N- cyclopenta -7- (2- methyl Pyridin-4-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (275mg, 0.727mmol, 36.7% yield), it is yellow solid, (R_fValue：0.4,10% methanol/DCM), LCMS (m/z): 364.29(M+H)⁺。

¹H NMR(400MHz,DMSO-d₆) δ ppm 10.30 (d, J=6.58Hz, 1H), 8.56 (d, J=5.04Hz, 1H), 7.67-7.52 (m, 4H), 5.41 (dd, J=5.81,2.96Hz, 1H), 4.14-4.06 (m, 1H), 3.21-3.02 (m, 2H), 2.99-2.85 (m, 2H), 2.54 (s, 3H), 2.17 (dd, J=9.76,3.84Hz, 1H), 2.03-1.79 (m, 3H), 1.64- 1.43(m,6H)。

Embodiment 176

Synthesize (4S)-N- (pyrazine -2- bases) -7- (1H- pyrazoles -5- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines with argon-degassed 20min And [2,3-b] [1,4] diaza- 5 (2H)-formamides (300mg, 0.947mmol), (1H- pyrazoles -5- bases) boric acid (127mg, 1.137mmol) and K₃PO₄Add in the solution of (402mg, 1.894mmol) in 1,4- dioxanes (9mL), water (3mL) Enter X-phos (45.2mg, 0.095mmol) and three (dibenzalacetone) two palladium (0) (43.4mg, 0.047mmol), use again Argon-degassed 10min.Reactant mixture is stirred into 4h at 100 DEG C, room temperature is subsequently cooled to.By reactant mixture through diatomite mistake Filter, then filtrate water dilutes and extracted (3 × 10mL) with EtOAc.Organic layer water, the aqueous salt solu-tion of merging, through sulphur Sour sodium is dried and evaporated, and obtains crude compound (TLC eluant, eluents：5%MeOH/DCM:R_f-0.3；UV activation).Rough chemical combination Thing is purified through column chromatography, is eluted using 100-200 silica gel mesh and with 2-3%MeOH/DCM, obtains pure (4S)-N- (pyrazine -2- Base) -7- (1H- pyrazoles -5- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first Acid amides (100mg, 0.280mmol, 29.5% yield), it is faint yellow solid, LCMS (m/z):349.2[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 14.58 (s, 1H), 12.8-11.0 (br, 1H), 9.50 (d, J=1.5Hz, 1H), 8.41 (dd, J=2.6,1.5Hz, 1H), 8.34 (d, J=2.6Hz, 1H), 7.69 (d, J=2.0Hz, 1H), 7.60 (d, J =7.9Hz, 1H), 7.41 (d, J=7.6Hz, 1H), 6.97 (d, J=17.7Hz, 1H), 5.63 (dd, J=6.0,3.2Hz, 1H), 3.33-3.14 (m, 3H), 3.03 (dd, J=12.1,3.3Hz, 1H), 2.35 (dddd, J=14.0,9.9,6.1, 4.1Hz,1H),2.17-2.02(m,1H)。

Embodiment 177

Synthesize 4- ((4S) -5- (pyrazine -2- bases carbamoyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 7- bases) pyridine -2- formic acid

Tripotassium phosphate (6.03g, 28.4mmol) is added into the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (3g, 9.47mmol), 4- (4,4,5,5- tetramethyls- 1,3- dioxolane -2- bases) pyridine -2- methyl formates (3.02g, 11.37mmol) are in 1,4- dioxanes (25mL) and water In the solution of stirring in (4.17mL).Reactant mixture is deaerated 15min, Pd is added₂(dba)₃(0.434g,0.474mmol) With X-phos (0.452g, 0.947mmol).Reactant mixture is deaerated 15min again, then 24hr are stirred at 100 DEG C.Will reaction Mixture is cooled to 28 DEG C, then distributes between water (50mL) and EtOAc (200mL).Water layer is with saturation citric acid soln With the solid of precipitation is filtered and dried.Crude solid is ground with first alcohol and water, 4- ((4S) -5- (pyrazine -2- base ammonia is obtained Base formoxyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 7- bases) pyridine -2- first Sour (272mg, 0.667mmol, 7.04% yield), it is pale solid.(TLC eluant, eluents：20%MeOH/DCM:R_f:0.1； UV activation), LCMS (m/z):404.1[M+H]⁺。

¹H NMR(DMSO-d₆,400MHz):δ 13.66 (s, 1H), 9.37 (d, J=0.8Hz, 1H), 8.87 (d, J= 4.8Hz, 1H), 8.62 (s, 1H), 8.42 (s, 1H), 8.38-8.34 (dd, J=8.4,2.4Hz, 2H), 7.887 (d, J=8Hz, 1H), 7.762 (d, J=8.4Hz, 1H) 5.53-5.51 (dd, J=5.6,2.8Hz, 1H), 3.31-3.23 (m, 1H), 3.15- 3.09 (m, 2H), 3.08-2.97 (dd, J=12,2.8Hz, 1H), 2.28-2.24 (m, 1H), 2.02-1.97 (m, 1H).

Embodiment 178

Synthesize (4S)-N- (6- picoline -3- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 30 DEG C to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaIn the solution of the stirring of (400mg, 1.585mmol) in THF (15mL) add triphosgene (282mg, 0.951mmol) and stirring 1h.Then Et is added at 30 DEG C₃N (1.105mL, 7.93mmol) and 6- picoline -3- amine (257mg, 2.378mmol), 16h is heated by reactant mixture at 65 DEG C.Reactant mixture is poured into and is used in combination in cold water (50mL) DCM extracts (2x50ml).The organic layer water of merging, salt water washing, it is dried over sodium sulfate and solvent is evaporated under reduced pressure, obtain thick production Thing.Crude mixture is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：0-15%MeOH/DCM), obtain (4S)- N- (6- picoline -3- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (320mg, 0.829mmol, 52% yield), it is pale solid (TLC:4%MeOH/ DCM,R_f:0.35), LCMS (m/z):387.25[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 12.94 (s, 1H), 8.66 (d, J=5.26Hz, 1H), 8.56 (d, J= 2.63Hz, 1H), 8.00 (dd, J=8.44,2.74Hz, 1H), 7.63 (d, J=8.14Hz, 1H), 7.57 (s, 1H), 7.50 (d, J=5.35Hz, 1H), 7.25-7.38 (m, 1H), 7.13 (d, J=8.55Hz, 1H), 5.70 (dd, J=5.92,3.29Hz, 1H), (s, the 3H) 2.03- of 3.10-3.35 (m, 3H), 3.02 (dd, J=12.06,3.29Hz, 1H), 2.58 (s, 3H), 2.48 2.22 (m, 1H) 2.34 (dddd, J=14.03,9.87,5.92,4.17Hz, 1H).

Embodiment 179

Synthesize (4S) -7- (2- (dimethylamino) pyridin-4-yl)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (300mg, 0.947mmol), K₃PO₄(603mg, 2.84mmol) in 1,4- dioxanes (30mL) and Added in the solution of degassing in water (0.3mL) x-phos (45.2mg, 0.095mmol) and acid chloride (II) (10.63mg, 0.047mmol).Then reactant mixture is heated into 12h at 120 DEG C.Solvent is evaporated under reduced pressure and is extracted (2x40ml) with DCM.Close And organic layer water, salt water washing, through anhydrous sodium sulfate drying and solvent is evaporated under reduced pressure, obtains crude product.Crude mixture Through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：3%MeOH/DCM), (4S) -7- (2- (dimethylaminos are obtained Base) pyridin-4-yl)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (210mg, 0.496mmol, 52.3% yield), it is pale solid (TLC:10%MeOH/EtOAc, R_f: 0.2), LCMS (m/z):403.31[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.56 (s, 1H), 9.54 (s, 1H), 8.25-8.11 (m, 3H), 7.62 (m, J=7.89Hz, 1H), 7.41 (d, J=7.89Hz, 1H), 7.19-7.01 (m, 2H), 5.72 (dd, J=5.81,3.18Hz, 1H), 3.31-3.14 (m, 9H), 3.02 (dd, J=12.17,3.18Hz, 1H), 2.50-2.23 (m, 1H), 2.10 (d, J= 7.02Hz,1H)。

Embodiment 180

Synthesize (4S) -7- (6- cyclopropyl pyridine -3- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature by potassium phosphate (402mg, 1.894mmol) and (6- cyclopropyl pyridine -3- bases) boric acid (185mg, 1.137mmol) add to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] Diaza- 5 (2H)-formamide (300mg, 0.947mmol) stirring in the mixture of n-butyl alcohol (6mL) and water (2.0mL) In the solution mixed.Purification for argon 30min is used, Pd is then added₂(dba)₃(43.4mg, 0.047mmol) and x-phos (45.2mg, 0.095mmol)), reactant mixture is stirred into 3h at 120 DEG C.Reactant mixture is reached room temperature, diluted with ethyl acetate (100mL), is washed with water (30mL) and salt solution (40mL).Organic layer is through Na₂SO₄It is dried and concentrated.By residue through quick post color Spectrum purifying (silica gel：100-200 mesh, 80% ethyl acetate/petroleum ether is used as eluant, eluent).By the substance migration ethanol of recovery and penta Alkane is recrystallized, and obtains (4S) -7- (6- cyclopropyl pyridine -3- bases)-N- (pyrazine -2- bases) -3,4- dihydro-Isosorbide-5-Nitrae-endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (130mg, 0.322mmol, 34.0% yield), it is canescence Solid (TLC eluant, eluents：100% ethyl acetate, R_f:0.2), LCMS (m/z):400.25[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 13.79 (s, 1H), 9.53 (d, J=1.5Hz, 1H), 9.01 (dd, J=2.4, 0.8Hz, 1H), 8.42-8.22 (m, 3H), 7.61 (d, J=8.0Hz, 1H), 7.45-7.19 (m, 2H), 5.70 (dd, J=5.9, 3.2Hz, 1H), 3.34-3.16 (m, 3H), 3.02 (dd, J=12.1,3.3Hz, 1H), 2.34 (dd, J=10.1,4.1Hz, 1H),2.19-2.02(m,2H),1.20-0.96(m,4H).

Embodiment 181

Synthesize (4S) -7- (piperidin-1-yl)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza- 5 (2H)-formamide

To the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (600mg, 1.894mmol) and piperidines (0.375mL, 3.79mmol) are at 1,4- dioxanes (10mL) Cs is added in the solution of middle degassing₂CO₃(1852mg, 5.68mmol), x-phos (361mg, 0.758mmol) and Pd (OAc)₂ (85mg,0.379mmol).Reactant mixture is stirred into 16h at 100 DEG C, by it down in cold water (20mL), extracted with ethyl acetate Take (2x50mL).The organic layer of merging is through anhydrous sodium sulfate drying and is concentrated under reduced pressure, and obtains crude product.Crude product is through quick post color Spectrum purifying (silica gel：100-200 mesh, eluant, eluent：5% methanol/DCM), obtain (4S) -7- (piperidin-1-yl)-N- (pyrazine -2- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (284mg, 0.762mmol, 40.2% yield), it is faint yellow solid (TLC:R_f:0.2, net EtOAc), LCMS (m/z):366.3[M+H ]⁺。

¹H NMR(400MHz,CDCl₃):δ 13.51 (s, 1H), 9.52 (d, J=1.5Hz, 1H), 8.29-8.19 (m, 1H), 8.18 (s, 1H), 7.33 (d, J=8.6Hz, 1H), 6.29 (d, J=8.6Hz, 1H), 5.61 (dd, J=6.1,3.3Hz, 1H), 3.55 (t, J=4.9Hz, 4H), 3.22 (dddd, J=12.2,8.6,3.7,2.3Hz, 1H), 3.16-3.07 (m, 2H), 2.90 (dd, J=11.8,3.3Hz, 1H), 2.34-2.22 (m, 1H), 2.00 (dddd, J=14.1,8.7,6.8,2.0Hz, 1H), 1.78-1.64(m,6H).

Embodiment 182

Synthesize (4S)-N- (1- methyl isophthalic acid H- pyrazole-3-yls) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 25 DEG C to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza(200mg,0.793mmol)、Et₃Stirrings of the N (0.331mL, 2.378mmol) in THF (10mL) Triphosgene (118mg, 0.396mmol) is added in solution, then reactant mixture is stirred at room temperature 30 minutes.Then 1- is added dropwise Solution of the methyl isophthalic acid H- pyrazoles -3- amine (231mg, 2.378mmol) in THF (2ml).By reactant mixture in 65 DEG C of stirrings 16h.Organic solvent is evaporated in vacuo and dilutes thick material (20ml) with DCM.Organic layer water (5mL), saturated brine (5mL) Washing, through anhydrous Na₂SO₄Dry and be evaporated in vacuo, obtain crude product.Crude mixture is through flash column chromatography (silica gel： 100-200 mesh, eluant, eluent：2%MeOH/DCM), obtain (4S)-N- (1- methyl isophthalic acid H- pyrazole-3-yls) -7- (2- picolines - 4- yls) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (185mg, 0.490mmol, 61.8% yield), it is white solid (TLC:5%MeOH/DCM, R_f:0.5), LCMS (m/z):376.3[M+ H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.37 (s, 1H), 8.61 (d, J=5.26Hz, 1H), 8.05-7.83 (m, 1H), 7.68-7.55 (m, 2H), 7.43 (d, J=7.89Hz, 1H), 7.35-7.17 (m, 1H), 6.63 (d, J=2.19Hz, 1H), 5.69 (dd, J=5.92,3.07Hz, 1H), 3.84 (s, 3H), 3.34-3.10 (m, 3H), 3.00 (dd, J=12.06, 3.29Hz, 1H), 2.75 (s, 3H), 2.32 (dddd, J=14.03,9.92,5.97,4.06Hz, 1H) 2.21-1.99 (m, 1H).

Embodiment 183

Synthesize (4S)-N- cyclobutyl -7- (2- picoline -4- bases) -3,4- dihydros -1,4- methylene-pyrido [2,3- B] [1,4] diaza- 5 (2H)-formamides

To (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysenes -1,4- that 30min is stirred at room temperature under a nitrogen Endo-methylene group pyrido [2,3-b] [1,4] diaza(400mg, 1.585mmol), triethylamine (1.105mL, 7.93mmol) With added in solution of the triphosgene (282mg, 0.951mmol) in tetrahydrofuran (20mL) cyclobutyl amine (225mg, 3.17mmol) the solution in THF (5mL).Reactant mixture is stirred into 16h at 65 DEG C, room temperature is subsequently cooled to.Reaction is mixed Compound is poured into frozen water and extracted (3 × 15mL) with EtOAc.Organic layer water, the aqueous salt solu-tion of merging, it is dry through sodium sulphate It is dry and evaporate, obtain crude compound (TLC eluant, eluents：5%MeOH/DCM:R_f-0.3；UV activation).Crude compound is passed through GRACE reversed-phase columns are purified, and are eluted with 30% (the 0.1%HCOOH aqueous solution)/MeOH, obtain (4S)-N- cyclobutyl -7- (2- Picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (369mg, 1.053mmol, 66.4% yield), it is faint yellow solid, LCMS (m/z):350.3[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 10.76-10.60 (m, 1H), 8.61 (dd, J=5.3,0.8Hz, 1H), 7.60 (dt, J=1.6,0.7Hz, 1H), 7.55 (d, J=7.8Hz, 1H), 7.49 (ddd, J=5.3,1.7,0.7Hz, 1H), 7.32 (d, J=7.9Hz, 1H), 5.62 (dd, J=6.0,3.2Hz, 1H), 4.52-4.36 (m, 1H), 3.28-3.13 (m, 2H), 3.07 (dt, J=11.9,2.1Hz, 1H), 2.94 (dd, J=12.0,3.3Hz, 1H), 2.65 (s, 3H), 2.51-2.39 (m, 2H), 2.26 (dddd, J=13.9,10.0,6.1,4.1Hz, 1H), 2.08-1.90 (m, 3H), 1.84-1.72 (m, 2H)

Embodiment 184

Synthesis (4S)-N- cyclopropyl -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza- 5 (2H)-formamides

Triphosgene (706mg, 2.378mmol) is added into (4S) -7- (2- picoline -4- bases) -2,3,4,5- four at 0 DEG C Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(600mg, 2.378mmol) and TEA (1.657mL, 11.89mmol) in the solution of the stirring in tetrahydrofuran (25mL).Reactant mixture is stirred into 1hr and cyclopropylamine is added (272mg, 4.76mmol), 16hr is stirred at 60 DEG C.Reactant mixture is cooled to 28 DEG C, then distribute in water (10mL) and Between EtOAc (25mL).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain rough Huang Color solid (TLC eluant, eluents：10%MeOH/EtOAc:R_f-0.2；UV activation).Thick material is purified through column chromatography, uses (100- 200 mesh) silica gel and eluted with 10%MeOH/EtOAc, pure (4S)-N- cyclopropyl -7- (2- picoline -4- bases) -3 is obtained, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (218mg, 0.638mmol, 26.8% yield), it is yellow solid, LCMS (m/z):336.24[M+H]⁺。

¹H NMR(CDCl₃,400MHz):δ 10.59 (s, 1H), 8.60 (d, J=5.4Hz, 1H), 7.68-7.50 (m, 2H), 7.42 (dd, J=5.2,1.7Hz, 1H), 7.38-7.21 (m, 1H), 5.66 (dd, J=5.9,3.2Hz, 1H), 3.36-3.12 (m, 2H), 3.07 (dt, J=12.0,2.1Hz, 1H), 3.01-2.82 (m, 2H), 2.66 (s, 3H), 2.27 (dddd, J= 13.8,9.9,6.0,4.0Hz, 1H), 2.02 (dt, J=14.6,7.3Hz, 1H), 0.91-0.79 (m, 2H), 0.69-0.51 (m, 2H).

Embodiment 185

Synthesize (4S) -7- (2- picoline -4- bases)-N- (pyridin-3-yl methyl) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Triphosgene (706mg, 2.378mmol) is added into (4S) -7- (2- picoline -4- bases) -2,3,4,5- four at 0 DEG C Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(600mg, 2.378mmol) and TEA (1.657mL, 11.89mmol) in the solution of the stirring in tetrahydrofuran (30mL).Reactant mixture is stirred into 1hr and pyridin-3-yl is added Methylamine (514mg, 4.76mmol), 16hr is stirred at 60 DEG C.Reactant mixture is cooled to 28 DEG C, then distributed at water (10mL) Between EtOAc (25mL).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain rough Yellow solid (TLC eluant, eluents：10%MeOH/EtOAc:R_f=0.2；UV activation).The inverted post purifying of crude compound (post：C18,40 μm) and eluted with 20% (0.1%HCOOH＆ water)/MeOH, obtain pure (4S) -7- (2- picolines -4- Base)-N- (pyridin-3-yl methyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5(2H)- Formamide (193mg, 0.498mmol, 20.93% yield), it is yellow solid, LCMS (m/z):387.32[M+H]⁺。

¹H NMR(CDCl₃,400MHz):δ 10.89 (t, J=5.7Hz, 1H), 8.75-8.60 (m, 1H), 8.56 (dd, J= 4.8,1.7Hz, 1H), 8.40 (d, J=5.3Hz, 1H), 7.72 (dt, J=7.9,2.0Hz, 1H), 7.57 (d, J=7.9Hz, 1H), 7.44-7.21 (m, 3H), 7.15 (dd, J=5.3,1.8Hz, 1H), 5.66 (dd, J=5.9,3.2Hz, 1H), 4.63 (d, J=5.4Hz, 2H), 3.37-3.15 (m, 2H), 3.11 (dt, J=12.2,2.1Hz, 1H), 2.98 (dd, J=12.0,3.3Hz, 1H), 2.49 (s, 3H), 2.39-2.23 (m, 1H), 2.05 (dt, J=14.6,7.8Hz, 1H).

Embodiment 186

Triphosgene (706mg, 2.378mmol) is added into (4S) -7- (2- picoline -4- bases) -2,3,4,5- four at 0 DEG C Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(600mg, 2.378mmol) and TEA (1.657mL, 11.89mmol) in the solution of the stirring in tetrahydrofuran (25mL).Reactant mixture is stirred into 1hr and pyridine -2- bases are added Methylamine (514mg, 4.76mmol), 16hr is stirred at 60 DEG C.Reactant mixture is cooled to 28 DEG C, then distributed at water (10mL) Between EtOAc (25mL).EtOAc layers of separation, then through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain thick thing Matter, it is yellow solid (TLC eluant, eluents：10%MeOH/EtOAc:R_f=0.2；UV activation).The inverted post of crude compound Purify (post：C18,40 μm) and eluted with 20% (0.1%HCOOH＆ water)/MeOH, obtain pure (4S) -7- (2- picolines - 4- yls)-N- (pyridine -2- ylmethyls) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (190mg, 0.492mmol, 20.67% yield), it is pale solid, LCMS (m/z):387.22[M+H ]⁺。

¹H NMR(CDCl₃,400MHz):δ 11.04 (t, J=5.4Hz, 1H), 8.45 (d, J=5.2Hz, 1H), 8.40 (dt, J=4.7,1.5Hz, 1H), 7.64 (td, J=7.7,1.9Hz, 1H), 7.60-7.49 (m, 2H), 7.44 (dd, J=5.3, 1.7Hz, 1H), 7.36 (d, J=7.8Hz, 1H), 7.33-7.22 (m, 1H), 7.22-7.10 (m, 1H), 5.68 (dd, J=6.0, 3.2Hz, 1H), 4.78 (d, J=5.0Hz, 2H), 3.34-3.04 (m, 3H), 2.97 (dd, J=12.0,3.3Hz, 1H), 2.49 (s, 3H), 2.28 (dddd, J=13.8,10.0,5.9,4.0Hz, 1H), 2.06 (dt, J=14.5,7.3Hz, 1H).

Embodiment 187

Synthesize (4S) -7- (2- picoline -4- bases)-N- (1,2,4- triazine -5- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

It is sub- to (4S) -7- (2- picoline -4- the bases) -2,3,4,5- tetrahydrochysene -1,4- bridges being stirred at room temperature under a nitrogen Picoline simultaneously [2,3-b] [1,4] diazaAdd in the solution of (0.250g, 0.991mmol) in tetrahydrofuran (30mL) Enter triethylamine (2.486mL, 17.83mmol) and triphosgene (0.294g, 0.991mmol).Then by reactant mixture in room temperature Stirring 30 minutes.1,2,4- triazine -5- amine (0.286g, 2.97mmol) is added in room temperature.Then by reactant mixture at 65 DEG C Stir 16hr.Reactant mixture is cooled to room temperature, solvent is completely distilled off out, then distributed in water (20mL) and EtOAc Between (50mL).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain thick material, it is palm fibre Color solid (TLC eluant, eluents：100%EtOAc:R_f-0.4；UV activation).Thick material is purified through column chromatography, uses (100-200 Mesh) silica gel and eluted with 100%EtOAc, obtain pure (4S) -7- (2- picoline -4- bases)-N- (1,2,4- triazine -5- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (0.044g, 0.117mmol, 11.76% yield), it is pale solid, LCMS (m/z):375.2[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 14.23 (s, 1H), 10.27 (d, J=2.0Hz, 1H), 9.27 (d, J= 2.1Hz, 1H), 8.65 (dd, J=5.3,0.8Hz, 1H), 7.96 (dt, J=1.9,0.7Hz, 1H), 7.73-7.61 (m, 2H), 7.54 (d, J=8.0Hz, 1H), 5.69 (dd, J=6.0,3.2Hz, 1H), 3.31-3.14 (m, 3H), 3.05 (dd, J=12.2, 3.3Hz, 1H), 2.73 (s, 3H), 2.38 (dddd, J=13.9,9.9,6.1,4.2Hz, 1H), 2.15-2.02 (m, 1H).

Embodiment 188

(4S)-N- cyclohexyl -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides

It is sub- to (4S) -7- (2- picoline -4- the bases) -2,3,4,5- tetrahydrochysene -1,4- bridges stirred under a nitrogen in room temperature Picoline simultaneously [2,3-b] [1,4] diazaThe solution of (500mg, 1.982mmol) in tetrahydrofuran (THF) (10mL) Middle addition triphosgene (294mg, 0.991mmol), is then stirred at room temperature 30min.To be added thereto triethylamine (1.381mL, 9.91mmol) with cyclo-hexylamine (255mg, 2.58mmol), reactant mixture is then stirred into 16hr at 60 DEG C.Reactant mixture Gone out with 2x15ml water quenchings and use 2x20ml ethyl acetate to extract, organic layer is dried over sodium sulfate and is concentrated under reduced pressure, and obtains roughization Compound.Purified on column chromatography is purified, and obtains pure compound (4S)-N- cyclohexyl -7- (2- picoline -4- bases) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (265mg, 0.701mmol, 35.4% yield), it is pale solid, (R_fValue：0.4,10% methanol/DCM), LCMS (m/z):378.36[M+H]⁺。

¹H NMR(400MHz,CDCl₃) δ ppm 10.36 (d, J=7.02Hz, 1H), 8.59 (d, J=5.26Hz, 1H), 7.56 (d, J=5.26Hz, 1H), 7.54-7.52 (m, 1H), 7.45 (dd, J=5.15,1.21Hz, 1H), 7.29 (d, J= 7.89Hz, 1H), 5.64 (dd, J=5.92,3.29Hz, 1H), 3.87-3.74 (m, 1H), 3.06-3.30 (m, 3H), 2.95 (dd, J=12.06,3.29Hz, 1H), 2.65 (s, 3H), 2.26 (dddd, J=14.00,9.89,5.92,4.17Hz, 1H), 2.14–1.98(m,3H),1.81-1.61(m,3H),1.45-1.14(m,5H)

Embodiment 189

Synthesis (4S)-N- (1H- imidazos [4,5-b] pyridine -5- bases) -7- (2- picoline -4- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaIn the solution of (600mg, 2.378mmol) in THF (30ml) add triphosgene (353mg, 1.189mmol), 1h is then stirred at room temperature.Then be sequentially added 1H- imidazos [4,5-b] pyridin-5-amine (415mg, 3.09mmol) and triethylamine (1.657mL, 11.89mmol) and by reactant mixture in seal pipe 70 DEG C heat 16h.Make Obtain reactant mixture and be cooled to room temperature, pour into saturation NaHCO₃Solution (150mL) is simultaneously extracted with ethyl acetate (3x50mL).Merge Organic layer through anhydrous Na₂SO₄Dry and be concentrated under reduced pressure, obtain crude product.Crude mixture is through flash column chromatography (silica gel： 100-200 mesh, eluant, eluent is used as using the gradient mixture in 1% ethanol/methylene), obtain (4S)-N- (1H- imidazos [4,5-b] pyridine -5- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] Diaza- 5 (2H)-formamides (160mg, 0.388mmol, 25%), it is faint yellow solid (TLC:5% methanol/DCM, R_f =0.3), LCMS (m/z):413.28[M+H]⁺。

¹H NMR(400MHz,DMSO-d6):δ 13.43 (s, 1H), 12.30-11.54 (m, 1H), 8.72 (d, J= 5.4Hz, 1H), 8.34 (s, 1H), 8.19-8.10 (m, 3H), 8.04 (d, J=8.7Hz, 1H), 7.80 (d, J=8.0Hz, 1H), 7.72 (d, J=8.0Hz, 1H), 5.55 (dd, J=6.0,3.1Hz, 1H), 3.12 (d, J=11.5Hz, 2H), 2.98 (d, J= 3.3Hz, 1H), 2.95 (d, J=3.3Hz, 1H), 2.70 (s, 3H), 2.25 (m, J=13.9,4.7Hz, 1H), 1.96 (dt, J= 14.3,7.4Hz,1H)。

Embodiment 190

Synthesize (4S)-N- (6- Calmazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Triethylamine (1.105mL, 7.93mmol) and triphosgene (235mg, 0.793mmol) are added into (4S) -7- at 25 DEG C (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(400mg, 1.585mmol) in the solution of the stirring in tetrahydrofuran (THF) (20mL), after 30min, 6- Calmazine -2- amine is added (359mg, 3.17mmol), is then heated to 70 DEG C, keeps 16h.Reactant mixture is set to reach room temperature, organic solvent passes through rotation Evaporation is removed.Residue diluted with water (50mL) is simultaneously extracted with ethyl acetate (3x60mL).The organic layer of merging salt water washing (60mL), through Na₂SO₄It is dried, filtered and concentrated, obtains crude compound.Crude compound is through flash column chromatography (silica gel: 100-200 mesh, 2%MeOH/DCM is used as eluant, eluent), obtain (4S)-N- (6- Calmazine -2- bases) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (210mg, 0.522mmol, 32.9% yield), it is faint yellow solid (TLC eluant, eluents：10%MeOH/DCM, R_f:0.4), LCMS (m/ z):392.26[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 13.82 (s, 1H), 9.48 (d, J=4.6Hz, 1H), 8.66 (d, J=5.3Hz, 1H), 8.15 (d, J=8.4Hz, 1H), 7.92 (d, J=2.2Hz, 1H), 7.71-7.40 (m, 2H), 7.26 (s, 1H), 5.75- 5.63 (m, 1H), 3.26 (dd, J=9.7,6.9Hz, 3H), 3.05 (d, J=3.3Hz, 1H), 2.75 (s, 3H), 2.36 (dddd, J=13.9,9.9,5.9,3.9Hz, 1H), 2.16-1.93 (m, 1H).

Embodiment 191

Synthesize (4S)-N- (pyrazine -2- bases) -7- (4- (trifluoromethyl) piperidin-1-yl) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (800mg, 2.53mmol) and 4- (trifluoromethyl) piperidines (580mg, 3.79mmol) are in 1,4- bis- Evil Cs is added in the solution of degassing in alkane (15mL)₂CO₃(2469mg, 7.58mmol), uses purification for argon 15min, is subsequently added into x- Phos (241mg, 0.505mmol), acid chloride (II) (56.7mg, 0.253mmol).By reactant mixture in seal pipe 100 DEG C of stirring 16h.So that reactant mixture is cooled to room temperature, pours into cold water (30mL) and be extracted with ethyl acetate (2x50mL).The organic layer of merging is concentrated under reduced pressure through anhydrous sodium sulfate drying, obtains crude product.Crude mixture is through quick post Chromatogram purification (silica gel：100-200 mesh, the gradient mixture using 2%MeOH/DCM is used as eluant, eluent), obtain (4S)-N- (pyrroles Piperazine -2- bases) -7- (4- (trifluoromethyl) piperidin-1-yl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine- 5 (2H)-formamides (255mg, 0.559mmol, 22.13% yield), it is pale solid (TLC:100% acetic acid Ethyl ester, R_f=0.3), LCMS (m/z):434.1[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 13.39 (s, 1H), 9.54 (d, J=1.5Hz, 1H), 8.25 (d, J=2.5Hz, 1H), 8.19 (dd, J=2.6,1.6Hz, 1H), 7.37 (d, J=8.6Hz, 1H), 6.32 (d, J=8.6Hz, 1H), 5.62 (dd, J=6.0,3.2Hz, 1H), 4.41-4.30 (m, 2H), 3.30-3.18 (m, 1H), 3.16-3.08 (m, 2H), 3.00-2.88 (m, 3H),2.37–2.19(m,2H),2.09–1.97(m,3H),1.75–1.61(m,2H).

Embodiment 192

Synthesize (4S) -7- (2- picoline -4- bases)-N- (2- (pyridin-3-yl) ethyl) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-carboxamide hydrochlorides

0 DEG C of diethyl ether solution (2mL, 4.00mmol) by 2.0M hydrochloric acid add to (4S) -7- (2- picoline -4- bases) - N- (2- (pyridin-3-yl) ethyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5(2H)- In formamide (250mg, 0.624mmol).Reactant mixture is stirred into 4h at 28 DEG C and concentrated, obtaining crude compound, (TLC is washed De- agent：5%MeOH/DCM:R_f-0.1；UV activation).Crude compound washs (2x5mL) with ether, obtains pure (4S) -7- (2- picoline -4- bases)-N- (2- (pyridin-3-yl) ethyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-carboxamide hydrochlorides (274mg, 0.606mmol, 97% yield), it is yellow solid, LCMS (m/ z):401.1[M+H]⁺。

¹H NMR(400MHz,CD₃OD):δ 8.89 (s, 2H), 8.72 (d, J=4.7Hz, 1H), 8.65 (d, J=6.7Hz, 1H), 8.44 (s, 1H), 8.32 (s, 1H), 8.17 (d, J=7.1Hz, 1H), 8.06-7.92 (m, 2H), 5.67 (d, J= 5.2Hz,1H),3.99-3.75(m,6H),3.32-3.20(m,2H),2.93(s,3H),2.72-2.53(m,1H),2.42- 2.37(s,1H)。

Embodiment 193

Synthesize ((4S) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] Diaza- 5 (2H)-yls) (3- methylpyrrolidin- 1- yls) ketone

It is sub- to (4S) -7- (2- picoline -4- the bases) -2,3,4,5- tetrahydrochysene -1,4- bridges stirred under a nitrogen in room temperature Picoline simultaneously [2,3-b] [1,4] diazaIn the solution of (1.5g, 5.94mmol) in tetrahydrofuran (THF) (20mL) Triphosgene (0.882g, 2.97mmol) is added, 30min is then stirred at room temperature.After 30 minutes, addition triethylamine (4.14mL, 29.7mmol) with 3- crassitudes (0.658g, 7.73mmol), reactant mixture is then stirred into 16hr at 60 DEG C.Reaction Mixture is gone out with 2x25ml water quenchings and uses 2x50ml ethyl acetate to extract, and organic layer is dried over sodium sulfate and is concentrated under reduced pressure, and obtains Crude compound.Purified on column chromatography purifies and carries out SFC, and the peak of separation is peak-I separation yields ((4S) -7- (2- methyl Pyridin-4-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-yls) (3- methyl pyrroles Cough up alkane -1- bases) ketone (210mg, 0.574mmol, 9.65% yield), it is Light brown solid, and peak-II separation yields ((4S) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-yl) (3- methylpyrrolidin- 1- yls) ketone (180mg, 0.494mmol, 6.79% yield), it is Light brown solid, (R_fValue：0.4,10% methanol/DCM), LCMS (m/z):364.32[M+H]⁺。

Peak -1

¹H NMR(400MHz,DMSO-d₆) δ ppm 8.50 (d, J=5.04Hz, 1H), 7.76 (d, J=0.66Hz, 1H), 7.68 (dd, J=5.26,1.10Hz, 1H), 7.52-7.48 (m, 1H), 7.47-7.38 (m, 1H), 4.40 (br d, J= 2.85Hz, 1H), 3.48 (br s, 3H), 3.16-2.98 (m, 4H), 2.89 (dd, J=11.62,3.29Hz, 1H), 2.53- 2.46 (m, 3H), 2.32-2.20 (m, 3H), 2.14 (br d, J=4.60Hz, 2H), 1.48 (dt, J=19.51,9.76Hz, 1H),1.12–0.98(m,2H)。

Peak-II

¹H NMR(400MHz,DMSO-d₆) δ ppm 8.48 (d, J=5.26Hz, 1H), 7.76 (s, 1H), 7.68 (dd, J= 5.26,1.32Hz,1H),7.40-7.54(m,2H),4.44(br s,1H),3.36-3.69(m,3H),2.99-3.18(m, 4H), 2.89 (dd, J=11.62,3.29Hz, 1H), 2.03-2.33 (m, 4H), 1.81-1.52 (m, 2H), 1.01 (br s, 2H)。

Embodiment 194

Synthesize (4S)-N- (5- picoline -3- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 30 DEG C to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaIn the solution of the stirring of (600mg, 2.378mmol) in THF (15mL) add triphosgene (423mg, 1.427mmol) and stirring 1h.Then Et is added₃N (1.657mL, 11.89mmol) and 5- picoline -3- amine (771mg, 7.13mmol) and by reactant mixture at 65 DEG C heat 16h.Reactant mixture is poured into cold water (50mL) and extracted with DCM (2x50ml).The organic layer water of merging, salt water washing, it is dried over sodium sulfate and solvent is evaporated under reduced pressure, obtain crude product.It is rough Mixture is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：0-15%MeOH/DCM), (4S)-N- (5- first is obtained Yl pyridines -3- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (300mg, 0.777mmol, 45% yield), it is yellow solid (TLC:10%MeOH/EtOAc, R_f:0.4), LCMS (m/z):387.36[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.00 (s, 1H), 8.66 (d, J=5.04Hz, 1H), 8.47 (s, 1H), 8.16 (s, 1H), 7.99 (s, 1H), 7.71-7.57 (m, 2H), 7.50 (d, J=5.26Hz, 1H), 7.38 (d, J=7.89Hz, 1H), 5.70 (dd, J=5.81,3.18Hz, 1H), 3.37-3.10 (m, 3H), 3.03 (dd, J=12.06,3.07Hz, 1H), 2.68(s,3H),2.45-2.25(m,4H),2.22–1.98(m,1H)。

Embodiment 195

Synthesize (4S)-N- (benzo [d] [1,3] dioxole -5- bases) -7- (2- picoline -4- bases) -3,4- two Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaIn the solution of (600mg, 2.378mmol) in THF (15mL) add triphosgene (353mg, 1.189mmol), 2h then is stirred at 27 DEG C, then adds benzo [d] [1,3] dioxole -5- amine at 27 DEG C (391mg, 2.85mmol) and N- ethyl-N-iospropyl propyl- 2- amine (307mg, 2.378mmol), is heated to 50 DEG C, keeps 16h. So that reactant mixture reaches room temperature and reactant mixture is poured into saturation NaHCO₃In solution and it is extracted with ethyl acetate (3x50mL).The organic layer of merging is through anhydrous Na₂SO₄Dry and be concentrated under reduced pressure, obtain crude product.Crude mixture is through quick post Chromatogram purification (silica gel：100-200 mesh, is eluted with 2% ethanol/methylene), obtain (4S)-N- (benzo [d] [1,3] dioxies Heterocyclic pentene -5- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine- 5 (2H)-formamides (270mg, 26.2% yield) (TLC:Eluant, eluent, 5% methanol/DCM；R_f=0.4), LCMS (m/ z):416.30[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 12.81 (s, 1H), 8.63 (d, J=5.26Hz, 1H), 7.64-7.59 (m, 2H), 7.49 (dd, J=5.15,1.43Hz, 1H), 7.37-7.31 (m, 2H), 6.85 (dd, J=8.33,2.19Hz, 1H), 6.75 (d, J=8.33Hz, 1H), 5.97-5.92 (m, 2H), 5.73-5.66 (m, 1H), 3.33-3.10 (m, 3H), 3.01 (dd, J=12.06,3.29Hz, 1H), 2.65 (s, 3H), 2.32 (dddd, J=14.11,9.84,5.86,4.17Hz, 1H), 2.09 (dt, J=14.03,6.80Hz, 1H).

Embodiment 196

Synthesize (4S) -7- (2- picoline -4- bases)-N- (2- (pyridine -2- bases) ethyl) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-carboxamide hydrochlorides

0 DEG C of diethyl ether solution (2mL, 4.00mmol) by 2.0M hydrochloric acid add to (4S) -7- (2- picoline -4- bases) - N- (2- (pyridine -2- bases) ethyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5(2H)- In formamide (300mg, 0.749mmol).Reactant mixture is stirred into 4h at 28 DEG C and concentrated, obtaining crude compound, (TLC is washed De- agent：5%MeOH/DCM:R_f-0.1；UV activation).Crude compound washs (3x5mL) with ether, obtains pure (4S) -7- (2- picoline -4- bases)-N- (2- (pyridine -2- bases) ethyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-carboxamide hydrochlorides (296mg, 0.675mmol, 90% yield), it is yellow solid, LCMS (m/ z)401.2[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ 9.88 (t, J=5.9Hz, 1H), 8.89 (d, J=6.1Hz, 1H), 8.66 (dd, J=5.6,1.7Hz, 1H), 8.40-8.35 (m, 1H), 8.31-8.29 (m, 1H), 8.14 (dd, J=6.2,1.9Hz, 1H), 8.08 (d, J=7.8Hz, 1H), 7.95 (dd, J=7.9,4.9Hz, 2H), 7.74 (t, J=6.7Hz, 1H), 5.40 (dd, J=5.6,2.9Hz, 1H), 3.97-3.78 (m, 2H), 3.58-3.26 (m, 6H), 2.84 (s, 3H), 2.32-2.25 (m, 1H), 2.19-2.03(m,1H)。

Embodiment 197

Synthesize (4S) -7- (piperazine -1- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to 4- ((4S) -5- (pyrazine -2- bases carbamoyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 7- bases) piperazine -1- t-butyl formates (359mg, 0.770mmol) are in THF (5mL) 2M HCl diethyl ether solution (1.924mL, 3.85mmol) is added in solution and reactant mixture is stirred into 4h at 35 DEG C, and will be anti- Answer mixture to be cooled to 0 DEG C, use NaHCO₃The aqueous solution neutralizes and uses CH₂Cl₂Extract (2x50mL).Organic layer water, the salt of merging Water washing, through anhydrous Na₂SO₄Dry and be concentrated under reduced pressure, obtain crude product.Crude mixture is through flash column chromatography (silica gel： 100-200 mesh, eluant, eluent：3%MeOH/DCM), obtain (4S) -7- (piperazine -1- bases)-N- (pyrazine -2- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(277mg, 0.741mmol, 96% are received -5 (2H)-formamides Rate), it is pale solid (TLC:Rf:0.3；5%MeOH/DCM), LCMS (m/z):367.2[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 13.41 (s, 1H), 9.53 (d, J=1.5Hz, 1H), 8.24 (d, J=2.5Hz, 1H), 8.17 (dd, J=2.5,1.5Hz, 1H), 7.38 (d, J=8.6Hz, 1H), 6.30 (d, J=8.6Hz, 1H), 5.62- 5.55 (m, 1H), 3.63 (t, J=5.2Hz, 4H), 3.26-3.17 (m, 1H), 3.17-3.06 (m, 6H), 2.92 (dd, J= 11.9,3.3Hz, 1H), 2.27 (dddd, J=13.7,9.9,6.2,3.8Hz, 1H), 2.07-1.96 (m, 2H)

Embodiment 198

Synthesize (4S) -7- (4- methyl piperidine -1- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (600mg, 1.894mmol), 4- methyl piperidines (376mg, 3.79mmol) are in Isosorbide-5-Nitrae-Er Evil In alkane (10mL) Cs is added in the solution of degassing₂CO₃(1852mg, 5.68mmol), x-phos (361mg, 0.758mmol) and Pd (OAc)₂(106mg,0.474mmol).Reactant mixture is stirred into 16h at 110 DEG C, pours into cold water (20mL), uses acetic acid second Ester extracts (2x50mL).The organic layer of merging is through anhydrous sodium sulfate drying and is concentrated under reduced pressure, and obtains crude product.Crude mixture is passed through Flash column chromatography (silica gel：100-200 mesh, eluant, eluent：4%MeOH/DCM), (4S) -7- (4- methyl piperidines -1- are obtained Base)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine (300mg, 0.775mmol, 40.9% yield), it is pale solid (TLC:R_f:0.4；Net EtOAc), LCMS (m/z): 380.3[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 13.51 (s, 1H), 9.52 (d, J=1.5Hz, 1H), 8.23 (dd, J=2.5, 0.4Hz, 1H), 8.20 (dd, J=2.5,1.5Hz, 1H), 7.33 (d, J=8.6Hz, 1H), 6.30 (d, J=8.7Hz, 1H), 5.61 (dd, J=6.1,3.2Hz, 1H), 4.20 (ddd, J=11.4,3.6,2.1Hz, 2H), 3.22 (dddd, J=12.3, 8.6,3.7,2.2Hz, 1H), 3.15-3.05 (m, 2H), 3.00-2.86 (m, 3H), 2.26 (dddd, J=13.8,9.9,6.2, 3.8Hz,1H),2.04-1.94(m,1H),1.84-1.72(m,2H),1.68-1.60(m,1H),1.33-1.18(m,2H), 0.99 (d, J=6.5Hz, 3H).

Embodiment 199

Synthesize (4S) -8- chloro- N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (1- ethyl -1H- pyrroles Azoles -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Under a nitrogen in room temperature to the chloro- N- of (4S) -8- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- Base) methoxyl group) pyridin-4-yl) -7- (1- ethyl -1H- pyrazoles -4- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza- 5 (2H)-formamides (450mg, 0.833mmol) add HCl in the solution in methanol (5mL) (3mL, 99mmol) and stir 2h.(5% methanol of TLC systems/DCM.Rf values:0.1), reactant mixture is concentrated, residue is 0 DEG C use saturation NaHCO₃It is basified.Gained solid is filtered, and is dried, is obtained crude compound, and it is used into triturated under ether (10mL), Obtain the chloro- N- of (4S) -8- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (1- ethyl -1H- pyrazoles -4- bases) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (280mg, 0.556mmol, 66.7% yield), it is pale solid.LCMS(m/z):500.26[M+H]⁺, Rt=1.74min.

¹H NMR(400MHz,DMSO-d6):δppm 12.66(s,1H),8.43(s,1H),8.10-7.92(m,2H), 7.73(s,1H),7.11-6.84(m,2H),5.52-5.33(m,1H),4.87(br s,1H),4.74-4.46(m,1H), 4.34-4.19 (m, 3H), 4.19-3.96 (m, 1H), 3.86-3.68 (m, 1H), 3.43 (br d, J=5.5Hz, 2H), 3.27- 3.15 (m, 1H), 3.14-2.99 (m, 2H), 2.99-2.72 (m, 1H), 2.36-2.08 (m, 1H), 1.93 (br dd, J=6.4, 13.8Hz, 1H), 1.46 (t, J=7.2Hz, 3H).

Embodiment 200

Synthesize (4S) -7- (2,6- lutidines -3- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

By K₃PO₄(535mg, 2.53mmol) adds to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamide (400mg, 1.263mmol) ＆ (2,6- lutidines -3- Base) stirring of the boric acid (248mg, 1.642mmol) in the dioxane of Isosorbide-5-Nitrae-(20mL) and water (1mL) solution in, then deaerate 15min simultaneously adds x-phos (60.2mg, 0.126mmol), is subsequently added into Pd₂(dba)₃(57.8mg, 0.063mmol), then will Reactant mixture heats 2h 45min at 90 DEG C, then allows to cool to room temperature, is filtered by Celite pad, is washed with ethyl acetate (10mL × 2) are washed, organic solvent is removed from filtrate decompression.Aqueous layer with ethyl acetate is extracted into (20mL × 2), with water (20mL × 2), salt solution (20mL) is washed, through Na₂SO₄Dry.Organic solvent is removed in vacuum, crude compound is obtained.Above-claimed cpd is through fast Fast column chromatography purifies (silica gel：100-200 mesh, is eluted with 80% ethyl acetate/hexane), obtain (4S) -7- (2,6- dimethyl pyrazoles Pyridine -3- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5(2H)- Formamide (330mg, 0.848mmol, 67.2% yield), it is pale solid (TLC:Eluant, eluent：100% ethyl acetate R_f: 0.3, UV activation), LCMS (m/z):388.28[M+H]⁺。

¹H-NMR(CDCl₃,400MHz):δ13.5(s,1H),9.45(s,1H),8.22-8.25(m,1H),8.18-8.20 (m, 1H), 7.77 (d, 1H, J=7.6Hz), 7.6 (d, 1H, J=8Hz), 7.08-7.14 (m, 2H), 5.68-5.72 (m, 1H), 3.18-3.34(m,3H),3.01–3.06(m,1H),2.65(s,3H),2.5(s,3H),2.31-2.39(m,1H),2.08- 2.15(m,1H)。

Embodiment 201

Synthesize (4S) -7- (2,6- lutidines -3- bases)-N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

By K₃PO₄(537mg, 2.53mmol) adds to (4S) -7- chloro- N- (pyridin-3-yl) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamide (400mg, 1.267mmol) ＆ (2,6- lutidines -3- Base) stirring of the boric acid (249mg, 1.647mmol) in the dioxane of Isosorbide-5-Nitrae-(15mL) and water (1mL) solution in, then deaerate 15min and successively addition dicyclohexyl (2', 4', 6'- triisopropyl-[1,1'- biphenyl] -2- bases) phosphine (60.4mg, 0.127mmol) and Pd₂(dba)₃(58.0mg, 0.063mmol), then heats 2h 45min at 100 DEG C.Reach reactant mixture To room temperature, filtered by Celite pad, (10mL × 2) are washed with ethyl acetate, the solvent of filtrate is removed under reduced pressure.By organic compound Thing is extracted with ethyl acetate (20mL × 2).The organic layer of merging is washed with water (20mL × 2), salt solution (20mL), through Na₂SO₄It is dry It is dry.Organic solvent is removed in vacuum, crude compound is obtained.Above-mentioned crude compound is through flash column chromatography (silica gel：100- 200 mesh, compound is eluted with 90% ethyl acetate/hexane), obtain (4S) -7- (2,6- lutidines -3- bases)-N- (pyrroles Pyridine -3- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (304mg, 0.777mmol, 61.3% yield), it is pale solid (TLC:Eluant, eluent：100% ethyl acetate R_f:0.4, UV activation ), LCMS (m/z):387.31[M+H]⁺。

¹H-NMR(CDCl₃,400MHz):δ13.13(s,1H),8.33–8.35(m,1H),8.24–8.26(m,1H), 8.10-8.14 (m, 1H), 7.6 (d, 1H, J=7.6Hz), 7.59 (d, 1H, J=8Hz), 7.19-7.24 (m, 1H), 7.15 (d, 1H, J=8Hz H), 7.05 (d, 1H, J=7.6Hz), 5.67-5.71 (m, 1H), 3.35-3.4 (m, 1H), 3.19-3.25 (m, 2H),3.1–3.36(m,1H),2.6(s,6H),2.29-2.38(m,1H),2.17–2.26(m,1H)。

Embodiment 202

Synthesize (4S) -7- (2- picoline -4- bases)-N- (6- methylthiazols simultaneously [3,2-b] [1,2,4] triazole -2- bases) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysenes -1,4- that 30min is stirred at room temperature under a nitrogen Endo-methylene group pyrido [2,3-b] [1,4] diaza(300mg, 1.189mmol), triethylamine (0.829mL, 5.94mmol) Be added dropwise in solution of the triphosgene (212mg, 0.713mmol) in tetrahydrofuran (10mL) 6- methylthiazols simultaneously [3,2-b] [1, 2,4] solution of the triazole -2- amine (367mg, 2.378mmol) in THF (2mL), continues 1min.By reactant mixture at 65 DEG C 16h is stirred, room temperature is subsequently cooled to.Reactant mixture is poured into water and extracted (3 × 10mL) with EtOAc.It is then combined with Organic layer water, aqueous salt solu-tion, it is dried over sodium sulfate, and evaporated, obtain crude compound (TLC eluant, eluents：10%MeOH/ DCM:R_f-0.1；UV activation).Crude compound inverted post purifying and with 90% (the 0.1%HCOOH aqueous solution)/MeOH Elution, obtains pure (4S) -7- (2- picoline -4- bases)-N- (6- methylthiazols simultaneously [3,2-b] [1,2,4] triazole -2- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (390mg, 0.892mmol, 75% yield), it is faint yellow solid, LCMS (m/z):433.2[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 13.88 (s, 1H), 8.63 (d, J=5.2Hz, 1H), 7.91 (d, J=1.9Hz, 1H), 7.69-7.52 (m, 2H), 7.46 (d, J=8.0Hz, 1H), 6.48 (d, J=1.6Hz, 1H), 5.75 (dd, J=5.9, 3.1Hz, 1H), 3.35-3.10 (m, 3H), 3.02 (dd, J=12.1,3.3Hz, 1H), 2.72 (s, 3H), 2.57 (d, J= 1.4Hz,3H),2.40-2.25(m,1H),2.19-2.05(m,1H)。

Embodiment 203

Synthesize (4S) -7- ((S) -3- (hydroxymethyl) morpholino)-N- (pyridin-3-yl) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature by Cs₂CO₃(5778mg, 17.74mmol) and (S)-morpholine -3- bases methanol (390mg, 3.33mmol) add To (4S) -7- chloro- N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 In the solution of stirring of (the 2H)-formamide (700mg, 2.217mmol) in the dioxane of Isosorbide-5-Nitrae-(10mL), purification for argon is then used 25min, then adds acid chloride (II) (49.8mg, 0.222mmol) and X-phos (211mg, 0.443mmol).Reaction is mixed Compound stirs 8h at 100 DEG C, then warms to room temperature reactant mixture.Organic solvent is removed by rotary evaporation, water is used Dilution (60mL) is simultaneously extracted with ethyl acetate (3x50mL).The organic layer of merging is with salt water washing (50mL), through Na₂SO₄Dry, Filter and concentrate, obtain crude residue.Crude residue purifies (silica gel through column chromatography：100-200 mesh, 2% methanol/dichloro Methane is used as eluant, eluent), (4S) -7- ((S) -3- (hydroxymethyl) morpholino)-N- (pyridin-3-yl) -3,4- dihydro -1 is obtained, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(275mg, 0.691mmol, 31.2% are received -5 (2H)-formamides Rate), it is pale solid (TLC eluant, eluents：10%MeOH/DCM R_f:0.4), LCMS (m/z):397.28[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 12.38 (s, 1H), 8.51 (d, J=2.6Hz, 1H), 8.36-8.24 (m, 1H), 8.16 (ddd, J=8.4,2.7,1.5Hz, 1H), 7.44-7.16 (m, 2H), 6.25 (d, J=8.6Hz, 1H), 5.63 (dd, J= 6.2,3.2Hz, 1H), 4.06 (d, J=7.4Hz, 2H), 3.81 (t, J=2.7Hz, 2H), 3.69-3.48 (m, 2H), 3.38- 3.03 (m, 4H), 2.90 (dd, J=11.8,3.3Hz, 1H), 2.26 (dd, J=10.1,3.9Hz, 1H), 2.11-1.87 (m, 1H), 1.26 (d, J=6.6Hz, 3H).

Embodiment 204

Synthesize (4S) -7- ((R) -3- methyl morpholine generations)-N- (pyridin-3-yl) -3,4- dihydros -1,4- methylene-pyridine And [2,3-b] [1,4] diaza- 5 (2H)-formamides

Successively (R) -3- methyl morpholines (320mg, 3.17mmol) and cesium carbonate (516mg, 1.583mmol) are added in room temperature To (4S) -7- chloro- N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 In the solution of stirring of (the 2H)-formamide (500mg, 1.583mmol) in the dioxane of Isosorbide-5-Nitrae-(8mL), then deaerate 30min. Then in room temperature by Pd (OAc)₂(71.1mg, 0.317mmol) and X-phos (755mg, 1.583mmol) add to reactant mixture And the 15min that deaerates again.Reactant mixture is stirred into 6h at 110 DEG C, room temperature is subsequently cooled to, (30mL) is diluted with water, second is used Acetoacetic ester extracts (2X50mL).The organic layer of merging is dried over sodium sulfate with salt water washing (20mL), filters and concentrates, and obtains Crude product.Crude mixture is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：2%MeOH/DCM), obtain (4S) -7- ((R) -3- methyl morpholine generations)-N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (100mg, 0.258mmol, 16.27% yield), it is pale solid (TLC: R_f0.4,10%MeOH/DCM), LCMS (m/z):381.26[M+H]⁺。

Embodiment 205

(4S)-N- (6- ((S) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (6- picoline -3- bases) -3 is synthesized, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

0 DEG C to (4S)-N- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine - 2- yls) -7- (6- picoline -3- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 4M HCl dioxane solutions are added in solution of (the 2H)-formamide (0.4g, 0.794mmol) in dichloromethane (10mL) (0.198g,1.589mmol).Then reactant mixture is stirred into 6h at 35 DEG C.By reactant mixture NaHCO₃Solution is neutralized And (2x30ml) is extracted with DCM.The organic layer of merging water (20mL), salt water washing, through anhydrous Na₂SO₄Dry and depressurize steaming Hair, obtains crude product.Crude compound is ground with ether (40mL) and pentane (20ml), obtains pure product (4S)-N- (6- ((S) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (6- picoline -3- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (0.21g, 0.453mmol, 65% yield), it is white solid (TLC:10%MeOH/ ethyl acetate, R_f:0.2), LCMS (m/z):464.27[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 12.99 (s, 1H), 9.33 (d, J=2.19Hz, 1H), 9.13 (s, 1H), 8.00 (d, J=7.78Hz, 2H), 7.97 (s, 1H), 7.64 (d, J=8.11Hz, 1H), 7.19-7.37 (m, 2H), 5.87 (br S, 1H), 5.73 (dd, J=5.92,3.29Hz, 1H), 4.74 (dd, J=10.96,5.70Hz, 1H), 4.40 (dd, J= 10.96,7.45Hz, 1H), 4.08-4.30 (m, 1H), 3.65-3.90 (m, 2H), 3.10-3.34 (m, 4H), 3.02 (dd, J= 12.28,3.29Hz, 1H), 2.62 (s, 3H), 2.36 (dddd, J=14.14,9.87,5.81,4.17Hz, 1H), 2.00-2.22 (m,1H)。

Embodiment 206

To the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (800mg, 2.53mmol) and 1- methyl piperazines (379mg, 3.79mmol) are in 1,4- dioxanes Cesium carbonate (2469mg, 7.58mmol), x-phos (241mg, 0.505mmol) and second are added in the solution of degassing in (15mL) Sour palladium (II) (56.7mg, 0.253mmol).Reactant mixture is heated into 16h at 100 DEG C and poured into cold water (45mL), second is used Acetoacetic ester extracts (2x100mL).The organic layer of merging is through anhydrous sodium sulfate drying and is concentrated under reduced pressure, and obtains crude product.It is rough mixed Compound is through flash column chromatography (silica gel：100-200 mesh, is eluted with 2%MeOH/DCM), obtain (4S) -7- (4- methyl piperazines Piperazine -1- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5(2H)- Formamide (268mg, 0.693mmol, 27.5% yield), it is yellow solid (TLC:100% ethyl acetate, R_f=0.23), LCMS(m/z):381.3[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δppm 13.46(s,2H),9.53(s,1H),8.29-8.12(m,2H),7.36 (d, J=8.55Hz, 1H), 6.30 (d, J=8.55Hz, 2H), 5.61 (dd, J=5.92,3.29Hz, 2H), 3.65-3.63 (m, 4H), 3.27-3.03 (m, 3H), 2.91 (dd, J=11.73,3.40Hz, 1H), 2.42-2.39 (m, 1H), 2.39 (s, 3H), 2.33-2.14-(m,1H),2.13-1.93(m,1H)。

Embodiment 207

Synthesize (4S) -7- (2- picoline -4- bases)-N- (5- (trifluoromethyl) pyrazine -2- bases) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In the molten of 25 DEG C of stirrings to 5- (trifluoromethyl) pyrazine -2- amine (1164mg, 7.13mmol) in THF (10mL) Triethylamine (1.989mL, 14.27mmol) and triphosgene (706mg, 2.378mmol) and stirring 1h are added in liquid.Then add (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza (600mg, 2.378mmol), then heats 15h in seal pipe at 100 DEG C.Reactant mixture is cooled to room temperature and acetic acid is used Ethyl ester extracts (2x50mL).The organic layer of merging water (20mL × 2), salt solution (10mL) washing, through anhydrous Na₂SO₄Dry and subtract Pressure concentration, obtains crude product.Crude product is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：2%MeOH/DCM), Obtain (4S) -7- (2- picoline -4- bases)-N- (5- (trifluoromethyl) pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (125mg, 0.283mmol, 32% yield), it is canescence Solid (TLC:Eluant, eluent：5% methanol/DCM, R_f:0.3), LCMS (m/z):442.24[M+H]⁺。

¹H-NMR(CDCl₃,400MHz):14.1(s,1H),9.7(s,1H),8.66-8.62(m,2H),8.0(s,1H), 7.69-7.64 (m, 2H), 7.5 (d, J=8Hz, 1H), 5.74-5.69 (m, 1H), 3.34-3.16 (m, 3H), 3.08-3.02 (m, 1H),2.75(s,3H),2.32-2.42(m,1H),2.14-2.06(m,1H)。

Embodiment 208

Synthesize (4S)-N- (pyrazine -2- bases) -7- (6- (trifluoromethyl) pyridin-3-yl) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

By the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (600mg, 1.894mmol), 5- (4,4,5,5- tetramethyls -1,3,2- dioxaborolans -2- Base) -2- (trifluoromethyl) pyridine (776mg, 2.84mmol) and K₃PO₄(804mg, 3.79mmol) is at 1,4- dioxanes (20mL) Deaerated 15 minutes in room temperature with the solution argon gas in water (3mL).By X-phos (90mg, 0.189mmol), and Pd₂(dba)₃ (87mg, 0.095mmol) adds to reactant mixture.Reactant mixture is deaerated again 15min and 90 DEG C stir 16h.Will reaction Mixture is cooled to 28 DEG C, is then filtered by Celite pad and evaporates filtrate, obtains crude residue.Crude residue water Dilution (20mL) is simultaneously extracted with ethyl acetate (2 × 80mL).The organic layer of merging is successively with water (20mL) and saline solution (10mL) is washed.Organic layer is dried over sodium sulfate, filters and evaporates filtrate, obtains brown solid.(TLC eluant, eluents：100% Ethyl acetate, R_f-0.3；UV activation).Crude compound is purified (neutral alumina) through column chromatography, product 45-50% second Acetoacetic ester/Hex, obtains (4S)-N- (pyrazine -2- bases) -7- (6- (trifluoromethyl) pyridin-3-yl) -3,4- dihydro -1, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(360mg, 0.824mmol, 43.5% are received -5 (2H)-formamides Rate), it is pale solid, LCMS (m/z):428.17[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 13.60 (s, 1H), 9.52 (d, J=1.1Hz, 1H), 9.28 (d, J=2.2Hz, 1H), 8.71 (dd, J=8.1,1.97Hz, 1H), 8.25-8.33 (m, 2H), 7.84 (d, J=8.3Hz, 1H), 7.67 (d, J= 7.9Hz, 1H), 7.5 (d, J=8Hz, 1H), 5.70 (dd, J=6.03,3.18Hz, 1H), 3.11-3.36 (m, 3H), 2.96- 3.11(m,1H),2.22-2.43(m,1H),1.97-2.21(m,1H)。

Embodiment 209

Synthesize (4S)-N- (isoquinolin -5- bases) -7- (6- picoline -3- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

It is sub- to (4S) -7- (6- picoline -3- the bases) -2,3,4,5- tetrahydrochysene -1,4- bridges being stirred at room temperature under a nitrogen Picoline simultaneously [2,3-b] [1,4] diazaThe solution of (0.750g, 2.97mmol) in tetrahydrofuran (THF) (40mL) Middle addition triethylamine (2.486mL, 17.83mmol) and triphosgene (0.882g, 2.97mmol).Reactant mixture is stirred in room temperature Mix 30 minutes, then add isoquinolin -5- amine (1.286g, 8.92mmol).Reactant mixture is stirred into 16hr at 65 DEG C, then Reactant mixture is cooled to room temperature and is concentrated under reduced pressure.By residue distribution between water (30mL) and dichloromethane (100mL). Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain thick material, it is that (TLC is washed brown solid De- agent：100%EtOAc:R_f-0.2；UV activation).Thick material is purified through column chromatography, using neutral alumina and with 100% 2 Chloromethanes is eluted, and obtains pure (4S)-N- (isoquinolin -5- bases) -7- (6- picoline -3- bases) -3,4- dihydro-Isosorbide-5-Nitraes-bridge sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (0.315g, 0.743mmol, 25.00% yield), its For pale solid, LCMS (m/z):423.3[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.21 (s, 1H), 9.20 (s, 1H), 8.88 (d, J=2.19Hz, 1H), 8.37 (d, J=7.23Hz, 1H), 7.94 (d, J=6.14Hz, 1H), 7.88 (dd, J=8.11,2.41Hz, 1H), 7.75 (d, J =8.11Hz, 1H), 7.61-7.69 (m, 2H), 7.47 (d, J=5.92Hz, 1H), 7.31 (d, J=7.89Hz, 1H), 7.08 (d, J=7.89Hz, 1H), 5.77 (dd, J=5.92,3.29Hz, 1H), 3.21-3.36 (m, 3H), 3.06 (dd, J=12.06, 3.07Hz, 1H), 2.59 (s, 3H), 2.32-2.43 (m, 1H), 2.17 (dt, J=14.03,7.02Hz, 1H.

Embodiment 210

Aza-tricycle [the 7.2.1.0 of the chloro- 5- of (9S) -4- (2- picoline -4- bases)-N- (pyridin-3-yl) -1,6,8- three { 2,7 }] 12 carbon -2,4,6- triolefin -8- formamides

At 0 DEG C (triethylamine is sequentially added to nicotinic acid in the solution of the stirring of 0.644g, 5.23mmol in THF (50mL) (3.6mL, 26.2mmol) and diphenyl phosphate azide (2.26mL, 10.46mmol).Make reaction solution that 2h is stirred at room temperature.Plus Enter the chloro- 7- of solid (4S) -8- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza(1.25g, 4.36mmol), 70 DEG C are heated to by reaction solution, are kept for 36 hours.Reaction solution distribution is existed EtOAc and H₂Between O.Organic layer is separated, through Na₂SO₄Dry, be concentrated in vacuo and be applied directly to silicagel column, use EtOAc/ EtOH(3:1) as eluant, eluent, (1g, 56%) is obtained, it is white solid.By gained solid Et₂O is ground, then vacuum Dry, LCMS (m/z):407.4[M+H]⁺。

¹H-NMR(400MHz,CHCl₃-d)δppm:d ppm 2.05-2.21(m,1H)2.28-2.50(m,1H)2.71(s, 3H) 2.97-3.46 (m, 4H) 5.69 (dd, J=5.81,3.03Hz, 1H) 7.14-7.32 (m, 1H) 7.43-7.58 (m, 1H) (d, J=2.27Hz, the 1H) 8.71 of 7.69 (s, 1H) 7.94-8.09 (m, 1H) 8.31 (dd, J=4.67,1.39Hz, 1H) 8.48 (d, J=5.05Hz, 1H) 12.64 (s, 1H).

Embodiment 211

Synthesize (4S) -7- (2,2- thebaine generations)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

Nitrine di(2-ethylhexyl)phosphate is added in 0 DEG C of solution to pyrazine -2- formic acid (350mg, 2.82mmol) in THF (15mL) Phenyl ester (1164mg, 4.23mmol) and DIPEA (2.463mL, 14.10mmol).Reactant mixture is stirred at 30 DEG C under a nitrogen Mix 2h.Add 2,2- dimethyl -4- ((4S) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas- 7- bases) morpholine (542mg, 1.974mmol) and by reactant mixture 65 DEG C stir 16h.Removal of solvent under reduced pressure.Thick material (20mL) is diluted with water and is extracted with ethyl acetate (2 × 30mL).The organic layer of merging is with water (10mL), and saturated brine solution is washed Wash.Organic layer filters through anhydrous sodium sulfate drying and evaporates filtrate, obtain crude product.Crude compound is purified through column chromatography (neutral alumina) product is eluted with 10% ethyl acetate/hexane.The fraction of collection is concentrated under reduced pressure, (4S) -7- (2,2- is obtained Thebaine generation)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (120mg, 0.303mmol, 10.74% yield), it is pale solid, LCMS (m/z):396.3[M+H ]⁺。

¹H-NMR(400MHz,CDCl₃):δ 13.40 (s, 1H), 9.56 (d, J=1.2Hz, 1H), 8.26 (d, J= 2.41Hz, 1H), 8.06-8.22 (m, 1H), 7.38 (d, J=8.8Hz, 1H), 6.28 (d, J=8.5Hz, 1H), 5.63 (dd, J =5.7,3.3Hz, 1H), 3.90 (t, J=5Hz, 2H), 3.47 (s, 2H), 3.41 (t, J=5Hz, 2H), 3.04-3.31 (m, 3H), 2.92 (dd, J=11.9,3.18Hz, 1H), 2.14-2.35 (m, 1H), 1.89-2.12 (m, 1H), 1.36 (s, 6H).

Embodiment 212

Synthesize (4S) -7- (6- methyl pyridazine -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

25 DEG C to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (400mg, 1.263mmol), 3- methyl -5- (4,4,5,5- tetramethyls -1,3,2- dioxies The miscellaneous amyl- 2- yls of boron heterocycle) pyridazine (306mg, 1.389mmol) and K₃PO₄(536mg, 2.53mmol) is at 1,4- dioxanes (15mL) With addition Pd (OAc) in the solution of the degassing in water (2mL)₂(14.18mg, 0.063mmol) and x-phos (60.2mg, 0.126mmol).Then reactant mixture is stirred into 8hr at 100 DEG C, reactant mixture is cooled to RT and is evaporated in vacuo Dioxane.Then it is diluted with water and is extracted (3x50mL) with DCM.The organic layer of merging is through anhydrous Na₂SO₄Dry and depressurize Solvent is removed, crude product is obtained.Crude mixture is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：2-3% MeOH/EtOAc), (4S) -7- (6- methyl pyridazine -4- bases)-N- (pyrazine -2- bases) -3,4- dihydros-Isosorbide-5-Nitrae-endo-methylene group is obtained Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (260mg, 0.689mmol, 54.6% yield), it is greyish white Color solid (TLC:10%MeOH/EtOAc, R_f:0.4), LCMS (m/z):375.25[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δppm 13.69(s,1H),9.62-9.53(s,2H),8.38-8.29(m,3H), 7.71 (d, J=8.11Hz, 1H), 7.59 (d, J=8.11Hz, 1H), 5.72 (dd, J=5.92,3.29Hz, 1H), 3.35- 3.15 (m, 2H), 3.12-3.00 (m, 2H), 2.90 (s, 3H), 2.48-2.25 (m, 1H), 2.11 (dt, J=14.03, 6.80Hz,1H)。

Embodiment 213

Synthesize (4S)-N- (6- fluorobenzene simultaneously [d] thiazol-2-yl) -7- (6- picoline -3- bases) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

It is sub- to (4S) -7- (6- picoline -3- the bases) -2,3,4,5- tetrahydrochysene -1,4- bridges being stirred at room temperature under a nitrogen Picoline simultaneously [2,3-b] [1,4] diazaThe solution of (0.750g, 2.97mmol) in tetrahydrofuran (THF) (40mL) Middle addition triethylamine (2.486mL, 17.83mmol) and triphosgene (0.882g, 2.97mmol).Reactant mixture is stirred in room temperature Mix 30 minutes, then adding 6- fluorobenzene in room temperature simultaneously [d] thiazole -2- amine (1.500g, 8.92mmol) and exists reactant mixture 65 DEG C of stirring 16hr, are subsequently cooled to room temperature and removal of solvent under reduced pressure.By the distribution of gained residue in water (30mL) and EtOAc Between (100mL).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain thick material, it is palm fibre Color solid (TLC eluant, eluents：100%EtOAc:R_f-0.2；UV activation).Thick material is purified through column chromatography, uses neutral alumina Aluminium, and use 100% dichloromethane eluent, obtains pure (4S)-N- (6- fluorobenzene simultaneously [d] thiazol-2-yl) -7- (6- methyl pyrroles Pyridine -3- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (0.313g, 0.700mmol, 23.53% yield), it is faint yellow solid, LCMS (m/z):447.2[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 14.74 (s, 1H), 9.06 (d, J=2.19Hz, 1H), 8.43 (dd, J= 8.11,2.41Hz, 1H), 7.75 (dd, J=8.88,4.71Hz, 1H), 7.66 (d, J=8.11Hz, 1H), 7.41-7.52 (m, 3H), 7.14 (td, J=8.99,2.63Hz, 1H), 5.68 (dd, J=5.92,3.07Hz, 1H), 3.17-3.33 (m, 3H), 3.04 (dd, J=12.17,3.18Hz, 1H), 2.68 (s, 3H), 2.32-2.43 (m, 1H), 2.12 (dt, J=14.20, 7.04Hz,1H)

Embodiment 214

Synthesize (4S) -7- (2- methylthiazol -5- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

Cesium fluoride (767mg, 5.05mmol) is added into the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (800mg, 2.53mmol), 2- methyl -5- (tributyls Stannyl) thiazole (1177mg, 3.03mmol) and tri-butyl phosphine (511mg, 2.53mmol) be at 1,4- dioxanes (40mL) In stirring solution in, deaerate 15min, adds acid chloride (II) (11.34mg, 0.051mmol) and cuprous iodide (I) (19.24mg,0.101mmol).Reactant mixture is deaerated 10min again, then 15hr are stirred at 100 DEG C.By reactant mixture 28 DEG C are cooled to, is then distributed between water (50mL) and EtOAc (125mL).Organic layer is separated, then through anhydrous Na₂SO₄It is dry It is dry, filtering.Evaporation filtrate obtains thick material.Thick material is purified through column chromatography, is used (100-200 mesh) silica gel, is used 5%MeOH/ EtOAc is eluted, and obtains (4S) -7- (2- methylthiazol -5- bases)-N- (pyrazine -2- bases) -3,4- dihydro-Isosorbide-5-Nitrae-endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (230mg, 0.600mmol, 23.77% yield), it is canescence Solid, LCMS (m/z):380.18[M+H]⁺。

¹H-NMR(CDCl₃,400MHz):δ 13.30 (s, 1H), 9.53 (d, J=1.6Hz, 1H), 8.43 (s, 1H), 8.36- 8.35 (dd, J=2.4,1.6Hz, 1H), 8.30 (d, J=2.8Hz 1H), 7.53 (d, J=8.0Hz, 1H), 7.18 (d, J= 8.0Hz, 1H), 5.68 (dd, J=6,3.2Hz, 1H), 3.28-3.22 (m, 2H), 3.20-3.13 (m, 1H), 3.02-2.98 (dd, J=12.4,3.6Hz, 1H), 2.78 (s, 3H), 2.35-2.32 (m, 1H), 2.10-2.04 (m, 1H)

Embodiment 215

Synthesize 4- ((4S) -5- (pyrazine -2- bases carbamoyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 7- bases) pyridine -2- methyl formates

Tripotassium phosphate (2.68g, 12.63mmol) is added into the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (2g, 6.31mmol), 4- (4,4,5,5- tetramethyls Base -1,3,2- dioxaborolan -2- bases) pyridine -2- methyl formates (1.994g, 7.58mmol) are in 1,4- dioxanes In the solution of stirring in (60mL).Reactant mixture is deaerated 15min.Add Pd₂(dba)₃(0.289g, 0.316mmol) and X-phos(0.301g,0.631mmol).Reactant mixture is deaerated 15min again, then 18hr are stirred at 100 DEG C.Reaction is mixed Compound is cooled to 28 DEG C, then distributes between water (50mL) and EtOAc (200mL).Organic layer and vacuum concentration are separated, is obtained Thick material (TLC eluant, eluents：5%MeOH/DCM:R_f=0.3；UV activation).Thick material is purified through column chromatography, uses (100- 200 mesh) silica gel, eluted with 10%MeOH/EtOAc, obtain 4- ((4S) -5- (pyrazine -2- bases carbamoyl) -2,3,4,5- Tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 7- bases) pyridine -2- methyl formates (780mg, 1.806mmol, 28.6% yield), it is pale solid, LCMS (m/z):418.23[M+H]⁺。

¹H NMR(CDCl₃,400MHz):δ 13.63 (s, 1H), 9.54 (d, J=1.2Hz, 1H), 8.91-8.89 (dd, J= 5.2,0.8Hz, 1H), 8.68-8.68 (dd, J=2,0.4Hz, 1H), 8.34-8.29 (m, 3H), 7.69 (d, J=8.0Hz, 1H), 7.59 (d, J=8.0Hz, 1H), 4.05 (s, 3H), 3.34-3.16 (m, 3H), 3.07-3.03 (dd, J=12.4, 3.2Hz,1H),2.41-2.33(m,1H),2.15-2.07(m,1H)。

Embodiment 216

Synthesize (4S) -7- (2- (difluoromethyl) pyridin-4-yl)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

28 DEG C by tripotassium phosphate (670mg, 3.16mmol) add to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (500mg, 1.579mmol), (2- (difluoros Methyl) pyridin-4-yl) stirring of the boric acid (328mg, 1.894mmol) in 1,4- dioxanes (10mL) and water (1.667mL) In solution.Reactant mixture is deaerated 15min, Pd is added₂(dba)₃(145mg, 0.158mmol), and X-phos (75mg, 0.158mmol).Reactant mixture is deaerated again 15min and by reactant mixture under microwave 100 DEG C stir 1hr.Will reaction Mixture is cooled to 28 DEG C, then distributes between water (10mL) and EtOAc (25mL).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain thick material, it is brown solid (TLC eluant, eluents：10%MeOH/EtOAc: R_f=0.3；UV activation).Thick material is purified through column chromatography, uses (100-200 mesh) silica gel, and use 5-10%MeOH/EtOAc Elution, obtains pure (4S) -7- (2- (difluoromethyl) pyridin-4-yl)-N- (pyrazine -2- bases) -3,4- dihydro-Isosorbide-5-Nitrae-bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (112mg, 0.273mmol, 17.27% yield), it is ash White solid, LCMS (m/z):410.17[M+H]⁺。

¹H NMR(CDCl₃,400MHz):δ 13.63 (s, 1H), 9.54 (s, 1H), 8.79 (d, J=5.2Hz, 1H), 8.40 (s, 1H), 8.32 (d, J=3.2Hz, 2H), 8.06 (dd, J=5.2,0.8Hz, 1H), 7.68 (d, J=7.6Hz, 1H), 7.54 (d, J=8Hz, 1H), 6.90-6.62 (t, J=55.2Hz, 1H), 5.73-5.71 (dd, J=5.6,2.8Hz, 1H), 3.33- 3.16 (m, 3H), 3.06-3.03 (dd, J=12.0,3.2Hz, 1H), 2.41-2.33 (m, 1H), 2.14-2.07 (m, 1H).

Embodiment 217

Synthesize (4S) -7- (the fluoro- 2- picolines -4- bases of 5-)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

28 DEG C by tripotassium phosphate (804mg, 3.79mmol) add to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (600mg, 1.894mmol), the fluoro- 2- of 5- Methyl -4- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) pyridine (674mg, 2.84mmol) is in 1,4- bis- In oxane (20mL) and the solution of the stirring in water (3.33mL).Reactant mixture is deaerated 15min, Pd is added₂(dba)₃ (173mg, 0.189mmol), and X-phos (90mg, 0.189mmol).Reactant mixture is deaerated 15min again, and reaction is mixed Compound stirs 1hr in microwave at 100 DEG C.Reactant mixture is cooled to 28 DEG C, then distributed in water (10mL) and EtOAc Between (25mL).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain thick material, it is palm fibre Color solid (TLC eluant, eluents：10%MeOH/EtOAc；R_f=0.3；UV activation).Crude product with methanol and ether washing, are obtained (4S) -7- (the fluoro- 2- picolines -4- bases of 5-)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza- 5 (2H)-formamides (317mg, 0.788mmol, 41.6% yield), it is pale solid, LCMS (m/z):392.20[M+H]⁺。

¹H NMR(CDCl₃,400MHz):δ 13.54 (s, 1H), 9.57 (d, J=1.6Hz, 1H), 8.45 (d, J=3.2Hz, 1H), 8.31 (d, J=2.8Hz, 1H), 8.27-8.26 (t, J=1.6Hz, 1H), 8.09 (d, J=6.8Hz, 1H), 7.66- 7.62 (t, J=8Hz, 2H), 5.73-5.71 (dd, J=6.0,3.2Hz, 1H), 3.34-3.17 (m, 3H), 3.06-3.02 (dd, J=12.0,3.2Hz, 1H), 2.71 (s, 3H), 2.40-2.33 (m, 1H), 2.13-2.06 (m, 1H).

Embodiment 218

Synthesize 4- ((4S) -5- (pyrazine -2- bases carbamoyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 7- bases) pyridine -2- methyl formates:

At 0 DEG C by NaBH₄(136mg, 3.59mmol) add to 4- ((4S) -5- (pyrazine -2- bases carbamoyl) -2,3, 4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 7- bases) pyridine -2- methyl formates (600mg, 1.437mmol) in the solution in THF (100mL).Reactant mixture is stirred into 8h at 28 DEG C.Solvent is passed through into rotary evaporation Remove, and by residue distribution between ethyl acetate (40mL) and water (20mL).Organic layer is separated, through anhydrous Na₂SO₄Dry And filter；Filter vacuum is evaporated, thick material is obtained.Thick material is purified by preparation HPLC, (4S) -7- (2- (hydroxyls are obtained Ylmethyl) pyridin-4-yl)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (135mg, 0.345mmol, 24.00%), it is pale solid, LCMS (m/z):390.22[M+H ]⁺。

¹H NMR(CDCl₃,400MHz):δ 13.66 (s, 1H), 9.60 (s, 1H), 8.68 (d, J=5.2Hz, 1H), 8.33 (s, 2H), 8.19 (s, 1H) 7.74-7.72 (dd, J=5.2,1.6Hz, 1H), 7.67 (d, J=8.0Hz, 1H), 7.52 (d, J= 8.0Hz, 1H), 5.73-5.70 (dd, J=6.0,3.6Hz, 1H), 4.93 (s, 2H), 3.31-3.16 (m, 3H), 3.06-3.02 (dd, J=12.4,3.2Hz, 1H), 2.38-2.35 (m, 1H), 2.12-2.08 (t, J=7.6Hz, 1H).

Embodiment 219

Synthesize (4S) -7- (6- cyanopyridine -3- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

By the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (600mg, 1.894mmol), 5- (4,4,5,5- tetramethyl -1,3,2- dioxaborolans -2- Base) pyridine -2- formonitrile HCNs (523mg, 2.273mmol) and K₃PO₄(804mg, 3.79mmol) is in 1,4- dioxanes (20mL), water Solution in (3mL) argon-degassed 20min, adds X-Phos (90mg, 0.189mmol), three (dibenzalacetone) two palladium (0) (87mg, 0.095mmol), uses argon-degassed 10min again.Reactant mixture is stirred into 16h at 100 DEG C, is subsequently cooled to Room temperature.Reactant mixture is filtered through diatomite, then filtrate water dilutes and extracted (3 × 15mL) with EtOAc.What is merged has Machine layer water, aqueous salt solu-tion, it is dried over sodium sulfate, and evaporated, obtain crude compound (TLC eluant, eluents：5%MeOH/ DCM:R_f-0.1；UV activation).Crude compound is purified through column chromatography, using neutral alumina and uses 30%EtOAc/ oil Ether is eluted, and obtains pure (4S) -7- (6- cyanopyridine -3- bases)-N- (pyrazine -2- bases) -3,4- dihydro-Isosorbide-5-Nitrae-endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (220mg, 0.568mmol, 30.0% yield), it is canescence Solid, LCMS (m/z):385.3[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.54 (s, 1H) 9.54 (d, J=1.32Hz, 1H) 9.34 (dd, J= 2.30,0.77Hz, 1H) 8.66 (dd, J=8.11,2.41Hz, 1H) 8.35-8.30 (m, 2H) 7.86 (dd, J=8.11, 0.88Hz, 1H) 7.70 (d, J=7.89Hz, 1H) 7.50 (d, J=8.11Hz, 1H) 5.71 (dd, J=5.92,3.29Hz, 1H) 3.35-3.14(m,3H)3.09-3.02(m,1H)2.43-2.32(m,1H)2.16-2.05(m,1H)。

Embodiment 220

Synthesize (4S)-N- (5- ethyl pyrazine -2- bases) -7- (6- picoline -3- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4S) -7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysenes -1,4- that 30min is stirred at room temperature under a nitrogen Endo-methylene group pyrido [2,3-b] [1,4] diaza(600mg, 2.378mmol), triphosgene (706mg, 2.378mmol) and 5- ethyl pyrazine -2- amine is added in solution of the triethylamine (1.989mL, 14.27mmol) in tetrahydrofuran (THF) (15mL) (879mg,7.13mmol).Reactant mixture is stirred into 16h at 70 DEG C, room temperature is subsequently cooled to, then falls reactant mixture Enter in water and extracted (3 × 15mL) with EtOAc.Organic layer water, the aqueous salt solu-tion of merging, it is dried over sodium sulfate and steam Hair, obtains crude compound (TLC eluant, eluents：5%MeOH/DCM:R_f-0.1；UV activation).Crude compound is pure through column chromatography Change, eluted using neutral alumina and with 30%EtOAc/ petroleum ethers, obtain pure (4S)-N- (5- ethyl pyrazine -2- bases) -7- (6- picoline -3- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine (383mg, 0.953mmol, 40.1% yield), LCMS (m/z):402.3[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.65 (s, 1H) 9.41 (d, J=1.53Hz, 1H) 9.11 (d, J= 2.19Hz, 1H) 8.41 (dd, J=8.22,2.52Hz, 1H) 8.22 (d, J=1.32Hz, 1H) 7.61 (d, J=7.89Hz, 1H) 7.40 (d, J=8.11Hz, 1H) 7.32 (d, J=8.11Hz, 1H) 5.70 (dd, J=6.03,3.18Hz, 1H) 3.34-3.13 (m, 3H) 3.02 (dd, J=12.06,3.29Hz, 1H) 2.83 (q, J=7.53Hz, 2H) 2.64 (s, 3H) 2.40-2.34 (m, 1H) 2.15-2.03 (m, 1H) 1.33 (t, J=7.56Hz, 3H).

Embodiment 221

Synthesize (4S)-N- (6- isopropoxy pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Successively triethylamine (1.326mL, 9.51mmol) and triphosgene (470mg, 1.585mmol) are added in room temperature (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza In the solution of the stirring of (400mg, 1.585mmol) in tetrahydrofuran (THF) (8mL) and stirring 30min, then by 6- isopropyls Epoxide pyrazine -2- amine (486mg, 3.17mmol) adds to reactant mixture, then stirs to 80 DEG C, keeps 16h.By reaction mixing Thing is cooled to room temperature, and (50mL) is diluted with water, and (2x40mL) is extracted with ethyl acetate, with salt water washing (30mL).Separation is organic Layer, it is dried over sodium sulfate, filter and concentrate.Residue is through purification by flash chromatography, using silica gel (100-200 mesh), and 2% methanol/ DCM, obtains (4S)-N- (6- isopropoxy pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro-Isosorbide-5-Nitrae-bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (60mg, 0.132mmol, 8.33% yield), it is greyish white Color solid (TLC:R_f:0.4,5% methanol/DCM), LCMS (m/z):432.27[M+H]⁺。

¹H NMR(400MHz,CDCl₃) δ ppm 1.24 (dd, J=13.15,6.14Hz, 6H) 2.09 (dt, J=14.14, 6.96Hz, 1H) 2.22-2.45 (m, 1H) 2.65 (s, 3H) 3.03 (dd, J=12.06,3.29Hz, 1H) 3.13-3.38 (m, 3H) 5.10 (dt, J=12.44,6.17Hz, 1H) 5.71 (dd, J=6.03,3.18Hz, 1H) 7.27-7.46 (m, 1H) 7.51-7.68 (s, the 1H) 13.04 of (m, 2H) 7.73 (dd, J=5.15,1.43Hz, 1H) 7.88 (s, 1H) 8.58 (d, J=5.26Hz, 1H) 9.03 (s,1H)。

Embodiment 222

Synthesize (4S) -7- (6- cyano group -5- picoline -3- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous(4,4,5,5- tetramethyl -1,3,2- dioxa boron is miscellaneous by -5 (2H)-formamides (500mg, 1.579mmol), 3- methyl -5- The amyl- 2- yls of ring) pyridine -2- formonitrile HCNs (501mg, 2.052mmol) and K₃PO₄(1.005g, 4.74mmol) is in 1,4- dioxanes X-phos (75mg, 0.158mmol) and Pd (OAc) is added in (20mL) and water (3mL) in the solution of degassing₂(17.72mg, 0.079mmol).Reactant mixture is deaerated again 15min and by reactant mixture 90 DEG C stir 16h.Reactant mixture is cold But to 28 DEG C, and filtered by Celite pad and evaporate filtrate, obtain residue.Residue diluted with water (20mL) is used in combination Ethyl acetate extracts (2 × 70mL).The organic layer of merging is successively washed with water (10mL) and saline solution (10mL).Organic layer is passed through Anhydrous sodium sulfate drying, filters and evaporates filtrate, obtain rough thing.(TLC eluant, eluents：100% ethyl acetate, R_f-0.3；UV Activation).Crude compound is purified through column chromatography, using neutral alumina, and product is eluted with 55-60% ethyl acetate/hexanes, Obtain pure (4S) -7- (6- cyano group -5- picoline -3- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (350mg, 0.835mmol, 52.9% yield), it is faint yellow Solid, LCMS (m/z):399.22[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 13.61 (s, 1H), 9.58 (d, J=1.5Hz, 1H), 9.07 (d, J=2.2Hz, 1H), 8.63-8.69 (m, 1H), 8.33 (d, J=2.8Hz, 1H), 8.28 (dd, J=1.4,2.6Hz, 1H), 7.69 (d, J= 8.1Hz, 1H), 7.52 (m, J=8.1Hz, 1H), 5.72 (dd, J=5.9,3.07Hz, 1H), 3.13-3.36 (m, 3H), 3.05 (dd, J=12.28,3.3Hz, 1H), 2.73 (S, 3H), 2.25-2.44 (m, 1H), 2.04-2.24 (m, 1H).

Embodiment 223

Synthesize (4S) -7- (2- methyl -6- (trifluoromethyl) pyridin-4-yl)-N- (pyrazine -2- bases) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous(4,4,5,5- tetramethyl -1,3,2- dioxa boron is miscellaneous by -5 (2H)-formamides (400mg, 1.263mmol), 2- methyl -4- The amyl- 2- yls of ring) -6- (trifluoromethyl) pyridine (471mg, 1.642mmol) and K₃PO₄(804mg, 3.79mmol) is in 1,4- bis- Evil X-phos (60.2mg, 0.126mmol) and Pd is added in alkane (20mL) and the solution of the degassing in water (3mL)₂(dba)₃ (57.8mg,0.063mmol).Reactant mixture is deaerated again 15min and by reactant mixture 90 DEG C stir 16h.Will reaction Mixture is cooled to 28 DEG C, and is filtered by Celite pad and evaporate filtrate, obtains residue.By residue diluted with water (20mL) and it is extracted with ethyl acetate (2 × 70mL).The organic layer of merging is successively washed with water (10mL) and saline solution (10mL) Wash.Organic layer is dried over sodium sulfate, filters and evaporates filtrate, obtains thick material (TLC eluant, eluents：100% ethyl acetate, R_f =0.4；UV activation).Crude compound purifies (neutral alumina) through column chromatography, by product with 45-50% ethyl acetate/oneself Alkane is eluted, and obtains (4S) -7- (2- methyl -6- (trifluoromethyl) pyridin-4-yl)-N- (pyrazine -2- bases) -3,4- dihydro-Isosorbide-5-Nitraes - Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (318mg, 0.719mmol, 56.9% yield), It is pale solid, LCMS (m/z):442.24[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 13.63 (s, 1H), 9.57 (d, J=1.3Hz, 1H), 8.22-8.40 (m, 2H), 8.14 (s, 1H), 8.11 (s, 1H), 7.69 (d, J=8.1Hz, 1H), 7.53 (d, J=7.9Hz, 1H), 5.73 (dd, J=5.9, 3.0Hz,1H),3.12-3.37(m,3H),2.95-3.12(m,1H),2.79(S,3H),2.25-2.46(m,1H),2.00- 2.21(m,1H)。

Embodiment 224

Synthesize (4S)-N- (pyridin-3-yl) -7- (6- (trifluoromethyl) pyridin-3-yl) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4S) -7- chloro- N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (500mg, 1.583mmol), 5- (4,4,5,5- tetramethyl -1,3,2- dioxaborolans -2- Base) -2- (trifluoromethyl) pyridine (562mg, 2.059mmol) and K₃PO₄(1008mg, 4.75mmol) is in 1,4- dioxanes X-phos (75mg, 0.158mmol) and Pd is added in (20mL) and water (3mL) in the solution of degassing₂(dba)₃(72.5mg, 0.079mmol).Reactant mixture is deaerated again 15min and by reactant mixture 90 DEG C stir 16h.Reactant mixture is cold But to 28 DEG C, and filtered by Celite pad and evaporate filtrate, obtain crude residue.Crude residue is diluted with water (20mL) And (2 × 70mL) is extracted with ethyl acetate.The organic layer of merging is successively washed with water (10mL) and saline solution (10mL).It is organic Layer is dried over sodium sulfate, filters and evaporates filtrate, obtains brown solid (TLC eluant, eluents：100% ethyl acetate, R_f-0.4； UV activation).Crude compound is purified through column chromatography, using neutral alumina, by product 35-40% ethyl acetate/hexanes Elution, obtains (4S)-N- (pyridin-3-yl) -7- (6- (trifluoromethyl) pyridin-3-yl) -3,4- dihydro-Isosorbide-5-Nitrae-endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (400mg, 0.924mmol, 58.3% yield), it is canescence Solid, LCMS (m/z):427.20[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 12.79 (s, 1H), 9.16 (d, J=1.9Hz, 1H), 8.54 (d, J=2.4Hz, 1H), 8.32 (dd, J=1.2,4.8Hz, 1H), 8.29 (dd, J=2,8.4Hz, 1H), 7.99-8.19 (m, 1H), 7.87 (d, J =8.3Hz, 1H), 7.69 (d, J=7.6Hz, 1H), 7.38 (d, J=7.9Hz, 1H), 7.28-7.30 (m, 1H), 5.71 (dd, J =5.8,3.2Hz, 1H), 3.12-3.36 (m, 3H), 2.94-3.12 (m, 1H), 2.25-2.43 (m, 1H), 2.00-2.23 (m, 1H)。

Embodiment 225

Synthesize (4S) -7- (2,5- lutidines -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides:

By Pd₂(dba)₃It is chloro- that (0.087g, 0.095mmol) and X-phos (0.018g, 0.047mmol) add to (4S) -7- N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (0.3g, 0.947mmol), 1- ethyls -4- (4,4,5,5- tetramethyls -1,3,2- dioxaborolan -2- bases) -1H- pyrazoles (0.316g, 1.421mmol) and potassium dihydrogen phosphate (0.258g, 1.894mmol) are at 1,4- dioxanes (5mL):In water (1mL) In the solution of degassing (argon gas).Reactant mixture is deaerated again 10min and 90 DEG C stir 15h.Reactant mixture is cooled to 28 DEG C, then filtered by Celite pad.Filtrate water (50mL) and ethyl acetate (50mL) dilution.Separation organic layer simultaneously uses water With salt water washing.Organic layer filters through anhydrous sodium sulfate drying and filtrate evaporation is obtained into crude compound (TLC eluant, eluents： 10%MeOH/EtOAc；R_f- 0.35UV activation).Crude compound is purified through column chromatography, using neutral alumina and uses 75% Ethyl acetate/hexane is eluted, and obtains (4S) -7- (1- ethyl -1H- pyrazoles -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(0.26g, 0.688mmol, 72.6% are received -5 (2H)-formamides Rate), it is pale solid, LCMS (m/z):377.30[M+H]⁺。

¹H NMR(CDCl₃,400MHz):δ 14.15 (s, 1H), 9.60 (d, J=1.53Hz, 1H), 8.56 (s, 1H), 8.22-8.30 (m, 2H), 8.07 (s, 1H), 7.50 (d, J=7.89Hz, 1H), 7.14 (d, J=7.89Hz, 1H), 5.66 (dd, J=6.03,3.18Hz, 1H), 4.28 (q, J=7.38Hz, 2H), 3.11-3.38 (m, 3H), 2.97-3.07 (m, 1H), 2.32 (dddd, J=14.03,9.98,6.03,3.95Hz, 1H), 1.97-2.13 (m, 1H), 1.60 (d, J=7.20Hz, 3H).

Embodiment 226

Synthesize (4S) -7- (6- (difluoro-methoxy) pyridin-3-yl)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

By Pd₂(dba₎₃It is chloro- that (0.087g, 0.095mmol) and X-phos (0.018g, 0.047mmol) add to (4S) -7- N- (pyrazine -2- bases) 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (0.3g, 0.947mmol), 2- (difluoro-methoxy) -5- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) Pyridine (0.385g, 1.421mmol) and potassium dihydrogen phosphate (0.258g, 1.894mmol) are at 1,4- dioxanes (5mL):Water (1mL) In the solution of middle degassing.Reactant mixture is deaerated again 10min and 90 DEG C stir 15h.Reactant mixture is cooled to 28 DEG C, Then filtered by Celite pad.Filtrate water (50mL) and ethyl acetate (50mL) dilution.Separate organic layer and with water and salt Water washing.Filtrate evaporation is obtained crude compound (TLC eluant, eluents by organic layer through anhydrous sodium sulfate drying, filtering：10% MeOH/EtOAc:R_f- 0.4UV activation).Crude compound is purified through column chromatography, using neutral alumina and uses 75% acetic acid Ethyl ester/Hex, obtains (4S) -7- (6- (difluoro-methoxy) pyridin-3-yl)-N- (pyrazine -2- bases) -3,4- dihydro -1, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(0.210g, 0.493mmol, 52.1% are received -5 (2H)-formamides Rate), it is pale solid, LCMS (m/z):426.22[M+H]⁺。

¹H NMR(CDCl₃,400MHz):δ 13.70 (s, 1H), 9.54 (d, J=1.53Hz, 1H), 8.81 (dd, J= 2.63,0.66Hz, 1H), 8.57 (dd, J=8.55,2.63Hz, 1H), 8.27-8.32 (m, 2H), 7.74 (s, 1H), 7.63 (d, J=8.11Hz, 1H), 7.37-7.75 (m, 1H), 7.06 (dd, J=8.55,0.66Hz, 1H), 5.70 (dd, J=5.92, 3.29Hz, 1H), 3.13-3.33 (m, 3H), 3.03 (dd, J=12.17,3.18Hz, 1H), 2.29-2.41 (m, 1H), 2.07- 2.13(m,1H)。

Embodiment 227

Synthesize (4S) -7- (6- picoline -3- bases)-N- (2- (((S)-tetrahydrofuran -3- bases) epoxide) pyrimidine -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (S) -2- ((tetrahydrofuran -3- bases) epoxide) pyrimidine -4- amine (862mg, 4.76mmol), triphosgene DIPEA (2.077mL, 11.89mmol) and (4S) -7- is added in the solution of (423mg, 1.427mmol) in THF (15mL) (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(600mg, 2.378mmol).Reactant mixture is stirred into 16h at 80 DEG C, then THF is evaporated under reduced pressure, residue diluted with water simultaneously uses DCM Extract (2x25mL).The organic layer of merging is through anhydrous Na₂SO₄Dry and be evaporated under reduced pressure, obtain crude compound.Use preparative HPLC purifies crude compound, obtains (4S) -7- (6- picoline -3- bases)-N- (2- (((S)-tetrahydrofuran -3- bases) oxygen Base) pyrimidine-4-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (195mg, 0.421mmol, 17.72% yield), it is white solid (TLC:10%MeOH/EtOAc, R_f:0.4), LCMS (m/ z):460.28[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.67 (s, 1H), 9.16 (d, J=2.19Hz, 1H), 8.38 (d, J= 5.70Hz, 1H), 8.34-8.07 (m, 1H), 7.80 (d, J=5.70Hz, 1H), 7.63 (d, J=7.89Hz, 1H), 7.39 (d, J =8.11Hz, 1H), 7.33-7.23 (m, 1H), 5.76-5.58 (m, 1H), 4.03-3.83 (m, 4H), 3.35-3.10 (m, 3H), 2.49-2.24(m,1H),2.23-2.00(m,3H)。

Embodiment 228

Synthesize (4S) -7- (2- methylpiperazine-1-yls)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to 3- methyl -4- ((4S) -5- (pyrazine -2- bases carbamoyl) -2,3,4,5- tetrahydrochysene -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 7- bases) piperazine -1- t-butyl formates (397mg, 0.826mmol) are in THF 2M HCl diethyl ether solution (2.065mL, 4.13mmol) is added in the solution of stirring in (5mL) and by reactant mixture 35 DEG C stirring 4h.By reactant mixture NaHCO₃The aqueous solution neutralizes and uses CH₂Cl₂Extract (3x50mL).The organic layer of merging is used Water, salt water washing, through Na₂SO₄Dry and be concentrated under reduced pressure, obtain crude product.Crude mixture is purified through preparation HPLC, is obtained (4S) -7- (2- methylpiperazine-1-yls)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (270mg, 0.710mmol, 85% yield), it is pale solid (TLC:5%MeOH/ DCM,Rf:0.3), LCMS (m/z):381[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δppm 13.36-13.49(m,1H),9.39(m,1H),8.41-8.22(m, 2H), 7.44-7.29 (m, 1H), 6.41 (dd, J=8.77,2.63Hz, 1H), 5.44 (dd, J=5.26,2.63Hz, 1H), 4.42-4.25(m,1H),3.93-3.68(m,1H),3.16–2.98(m,4H),2.90-2.73(m,3H),2.69-2.53(m, 1H), 2.37-2.27 (m, 2H), 2.26-2.10 (m, 1H), 1.99-1.79 (m, 1H), 1.12 (dd, J=6.58,3.95Hz, 3H)。

Embodiment 229

Synthesize (4S) -7- (3- methylpiperazine-1-yls)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature in a hydrogen atmosphere (gasbag pressure) to (4S) -7- (4- benzyl -3- methylpiperazine-1-yls)-N- (pyrroles Piperazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (450mg, Acetic acid (0.027mL, 0.478mmol) and 10% hydroxide 0.956mmol) are added in the solution of the stirring in methanol (10mL) Reactant mixture is simultaneously stirred 4h by palladium/charcoal (67.1mg, 0.096mmol) at 35 DEG C.After the completion of reaction, by reactant mixture through silicon Diatomaceous earth pad is filtered, and filter vacuum is evaporated, crude product is obtained.Crude mixture is purified through preparation HPLC, obtains (4S) -7- (3- methylpiperazine-1-yls)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (280mg, 0.736mmol, 76% yield), it is Light brown solid (TLC:Rf:20%MeOH/DCM, 0.4), LCMS (m/z):381.3[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.43 (s, 1H), 9.55 (d, J=1.53Hz, 1H), 8.24 (d, J= 2.63Hz, 1H), 8.09-8.20 (m, 2H), 7.36 (d, J=8.55Hz, 1H), 6.30 (d, J=8.55Hz, 1H), 5.62 (dd, J=5.92,3.07Hz, 1H), 4.20 (d, J=12.06Hz, 1H), 3.95 (d, J=11.84Hz, 1H), 3.08-2.87 (m, 4H) 3.25-3.08 (m, 4H), 2.57 (ddd, J=12.33,10.25,2.19Hz, 1H), 2.27 (dddd, J=13.92, 9.98,6.14,3.73Hz, 1H), 2.00 (dt, J=14.41,7.37Hz, 1H), 1.21 (d, J=6.14Hz, 3H).

Embodiment 230 and embodiment 231

(4S)-N- (pyrazine -2- bases) -7- ((±) -2- (trifluoromethyl) morpholino) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (2g, 6.31mmol) and 2- (trifluoromethyl) morpholine (1.469g, 9.47mmol) are in 1,4- dioxanes Cesium carbonate (6.17g, 18.94mmol), x-phos (1.204g, 2.53mmol) and acetic acid are added in (30mL) in the solution of degassing Palladium (II) (0.284g, 1.263mmol).Reactant mixture is stirred into 16h in seal pipe at 100 DEG C.Reactant mixture is fallen Enter cold water (80mL) and be extracted with ethyl acetate (3x100mL).The organic layer of merging is through anhydrous sodium sulfate drying and is concentrated under reduced pressure, Obtain crude product.Crude mixture is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：2%MeOH/CH₂Cl₂), obtain To (4S)-N- (pyrazine -2- bases) -7- ((±) -2- (trifluoromethyl) morpholino) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3-b] [1,4] diazaThe mixture of the enantiomter of -5 (2H)-formamides.By the chiral preparative SFC of mixture (the condition of separation：(cellulose -2 (250 × 30) mm, 55.0% (100%MeOH), 90.0g/min, 100.0 bars, 42nm, 8.5min)), peak-I (most fast eluates are obtained：320mg, 0.736mmol, 42% yield), and peak-II (most slow eluates： 305mg, 0.701mmol, 40.5% yield), it is pale solid (TLC：Net EtOAc, R_f:- 0.21), LCMS (m/z): 436.25[M+H]⁺。

The peak most eluted soon:¹H NMR(400MHz,CDCl₃):δ ppm 13.19 (s, 1H), 9.52 (s, 1H), 8.25 (d, J =2.41Hz, 1H), 8.21-8.09 (m, 1H), 7.43 (d, J=8.33Hz, 1H), 6.35 (d, J=8.55Hz, 1H), 5.64 (dd, J=6.03,3.18Hz, 1H), 4.21 (dd, J=11.84,1.97Hz, 1H), 4.15-4.00 (m, 1H), 4.00-3.77 (m, 3H), 3.26-3.04 (m, 5H), 3.04-2.82 (m, 1H), 2.28 (dddd, J=13.95,10.00,6.19,3.84Hz, 1H),2.07–1.92(m,1H)。

The peak most eluted slowly:¹H NMR(400MHz,CDCl₃):δ ppm 13.20 (s, 1H), 9.52 (d, J=1.53Hz, 1H), 8.26 (d, J=2.41Hz, 1H), 8.16 (dd, J=2.41,1.53Hz, 1H), 7.43 (d, J=8.55Hz, 1H), 6.35 (d, J=8.55Hz, 1H), 5.63 (dd, J=6.03,3.18Hz, 1H), 4.21 (d, J=12.06Hz, 1H), 4.08 (ddd, J =10.69,6.19,2.85Hz, 1H), 4.00-3.92 (m, 2H), 3.84 (td, J=11.56,2.96Hz, 1H), 3.26-3.07 (m,5H),3.07–2.82(m,1H),2.43-2.24(m,1H),2.21–1.95(m,1H)。

Embodiment 232

Synthesize ((R) -2- methyl morpholine generations) ((4S) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-yls) methanone hvdrochloric acid salt

1.0M hydrochloric acid dioxane solutions (1.6mL) are added into (R) -2- methyl morpholine generations at 0 DEG C) ((4S) -7- (2- first Yl pyridines -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-yls) ketone In the solution of the stirring of (0.21g, 0.544mmol) in methanol (5mL).Reactant mixture is evaporated, crude compound is obtained. Crude compound obtains pure ((R) -2- methyl morpholine generations) ((4S) -7- (2- picolines -4- with triturated under ether (3x5mL) Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-yls) methanone hvdrochloric acid salt (105mg, 0.253mmol, 45% yield), it is yellow solid, LCMS (m/z):380.25[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ 8.86 (br d, J=5.92Hz, 1H), 8.41 (br s, 1H), 8.28 (br s,1H),7.82-8.07(m,2H),4.64(br s,1H),3.23-4.11(m,9H),2.99-3.14(m,1H),2.53-2.87 (m,4H),2.19-2.43(m,2H),0.89-1.15(m,3H)

Embodiment 233

Synthesis (4S) -7- (1- methyl -2- oxo -1,2- dihydropyridine -4- bases)-N- (pyridin-3-yl) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature to (4S) -7- chloro- N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (400mg, 1.267mmol), potassium phosphate (537mg, 2.53mmol) and (1- methyl -2- oxygen Generation -1,2- dihydropyridine -4- bases) boric acid (291mg, 1.900mmol) is in 1,4- dioxanes (9mL) and the mixture of water (1mL) In degassing solution in add X-Phos (60.4mg, 0.127mmol) and Pd (OAc)₂(14.22mg, 0.063mmol), so Afterwards 16h is heated at 100 DEG C.Reactant mixture is set to reach room temperature, organic solvent is removed and is diluted with water by rotary evaporation (50mL), is extracted with ethyl acetate (3x50mL).The organic layer of merging is with salt water washing (50mL), through anhydrous Na₂SO₄Dry simultaneously Concentration, obtains crude product.Crude product is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：03%MeOH/DCM), obtain To (4S) -7- (1- methyl -2- oxo -1,2- dihydropyridine -4- bases)-N- (pyridin-3-yl) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (150mg, 0.383mmol, 30.2% yield), it is ash White solid (TLC:R_f0.4,10%MeOH/DCM), LCMS (m/z):389.22[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm12.92 (s, 1H), 8.57 (d, J=2.63Hz, 1H), 8.06-8.32 (m, 2H), 7.62 (d, J=7.89Hz, 1H), 7.48 (d, J=7.02Hz, 1H), 7.25-7.32 (m, 2H), 6.95 (d, J= 1.75Hz, 1H), 6.63 (dd, J=7.02,1.97Hz, 1H), 5.68 (dd, J=5.92,3.29Hz, 1H), 3.62 (s, 3H), 3.19-3.39 (m, 2H), 3.15 (d, J=12.06Hz, 1H), 2.84-3.09 (m, 1H), 2.19-2.39 (m, 1H), 2.05- 2.19(m,1H)。

Embodiment 234

Synthesize ((S) -2- methyl morpholine generations) ((4S) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-yls) methanone hvdrochloric acid salt

1.0M hydrochloric acid dioxane solutions (1.74mL) are added into (S) -2- methyl morpholine generations at 0 DEG C) (4S) -7- (2- first Yl pyridines -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-yls) ketone (0.22g, 0.580mmol) is in methanol (5mL) in the solution of stirring.Reactant mixture is stirred into 4h at 28 DEG C.Reaction is mixed Compound evaporates, and obtains crude compound.Crude compound obtains pure ((S) -2- methyl morpholines with triturated under ether (3x5mL) Generation) ((4S) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-yls) methanone hvdrochloric acid salt (110mg, 0.265mmol, 45% yield), it is yellow solid, LCMS (m/z):380.25[M +H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ 8.86 (d, J=6.36Hz, 1H), 8.38-8.45 (m, 1H), 8.24-8.34 (m, 1H), 7.86-7.99 (m, 2H), 4.57-4.64 (m, 1H), 4.07 (brd, J=10.30Hz, 1H), 3.35-3.85 (m, 10H), 3.07 (br t, J=12.06Hz, 1H), 2.81 (s, 4H), 2.39 (s, 2H), 0.96-1.17 (m, 3H).

Embodiment 235

Synthesize (4S)-N-7- two (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaIn the solution of the stirring of (0.8g, 3.17mmol) in THF (15mL) add triphosgene (0.565g, 1.902mmol) and stirring 30min.Then 2- picoline -4- amine (0.686g, 6.34mmol) and DIPEA are added in room temperature (1.661mL,9.51mmol).Reactant mixture is heated into 15h at 80 DEG C.THF is evaporated under reduced pressure and (15mL) is diluted with water simultaneously Extracted with DCM (2x20ml).The organic layer of merging is through anhydrous Na₂SO₄Dry and be evaporated under reduced pressure, obtain crude compound.Roughization Compound obtains (4S)-N-7- two (2- picoline -4- bases) -3,4- dihydro-Isosorbide-5-Nitrae-endo-methylene group pyrroles through flash column chromatography Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (120mg, 0.31mmol, 22%), it is white solid (TLC: 10%MeOH/EtOAc, R_f:0.4), LCMS (m/z):387.28[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.15 (s, 1H), 8.68 (d, J=5.26Hz, 1H), 8.36 (d, J= 5.48Hz, 1H), 7.70-7.56 (m, 1H), 7.56-7.47 (m, 1H), 7.34-7.42 (m, 2H), 7.31-7.25 (m, 2H), 5.68 (dd, J=5.92,3.07Hz, 1H), 3.20-3.11 (m, 3H), 3.03 (dd, J=12.06,3.29Hz, 1H), 2.63 (s,3H),2.52(s,3H),2.43-2.25(m,1H),2.15-2.03(m,1H)。

Embodiment 236

Synthesize (4S) -7- (4,6- lutidines -3- bases)-N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature to (4S) -7- chloro- N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (400mg, 1.267mmol), 2,4- dimethyl -5- (4,4,5,5- tetramethyls -1,3,2- Dioxaborolan -2- bases) pyridine (591mg, 2.53mmol) and Cs₂CO₃(1238mg, 3.80mmol) is in 1,4- dioxanes Pd (OAc) is added in the solution deaerated in the mixture of (9mL) and water (1mL)₂(14.22mg, 0.063mmol) and X-Phos (60.4mg, 0.127mmol), then heats 16h at 100 DEG C.Reactant mixture is set to reach room temperature, organic solvent steams by rotating Hair is removed and is diluted with water (50mL), is extracted with ethyl acetate (3x50mL).The organic layer of merging salt water washing (50mL), warp Anhydrous Na₂SO₄It is dried, filtered and concentrated, obtains crude product.Crude mixture is through flash column chromatography (silica gel：100-200 Mesh, eluant, eluent：3%MeOH/DCM), (4S) -7- (4,6- lutidines -3- bases)-N- (pyridin-3-yl) -3,4- bis- is obtained Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (115mg, 0.295mmol, 23.30% yield), it is pale solid (TLC:R_f:0.4,10%MeOH/DCM), LCMS (m/z):387.25[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.07 (br s, 1H), 8.51 (s, 1H), 8.24 (brd, J= 4.17Hz, 1H), 8.18-7.97 (m, 1H), 7.61 (br d, J=7.67Hz, 1H), 7.34-7.16 (m, 2H), 7.13 (m, 1H), 5.68 (br d, J=2.19Hz, 1H), 3.38-3.11 (m, 3H), 3.11-2.89 (m, 1H), 2.60 (s, 3H), 2.36 (s,4H)2.30-2.03(m,1H),1.36-1.12-(m,1H)。

Embodiment 237

Synthesis (4S) -7- (6- picoline -3- bases)-N- neopentyl -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (4S) -7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- methylene benzos [b] [1,4] DiazaIn the solution of the stirring of (250mg, 0.991mmol) in THF (10mL) add triphosgene (176mg, 0.594mmol) and stirring 30min.Then by DIPEA (0.519mL, 2.97mmol) and 2,2- dimethyl propylene -1- amine (130mg, Above-mentioned reactant mixture 1.486mmol) is added to, then 16h is stirred at 80 DEG C.Reactant mixture is warmed to room temperature, it is dilute with water Release (60mL) and be extracted with ethyl acetate (3x50mL).The organic layer of merging is with salt water washing (60mL), through anhydrous Na₂SO₄It is dry It is dry, filter and concentrate, obtain crude product.Thick material is through flash column chromatography (silica gel 100-200 mesh, eluant, eluent：2%MeOH/ DCM), (4S) -7- (6- picoline -3- bases)-N- neopentyls -3,4- dihydro-Isosorbide-5-Nitrae-endo-methylene group pyrido [2,3-b] is obtained [1,4] diaza- 5 (2H)-formamides (163mg, 0.441mmol, 44.5% yield), it is faint yellow solid (TLC:R_f: 0.4,10%MeOH/DCM), LCMS (m/z):366.29[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 10.45 (br s, 1H), 8.88 (s, 1H), 7.93 (br d, J= 7.89Hz, 1H), 7.54 (d, J=7.89Hz, 1H), 7.34-7.11 (m, 2H), 5.77-5.51 (m, 1H), 3.33-3.14 (m, 5H), 2.95 (brdd, J=11.84,3.07Hz, 1H), 2.61 (s, 3H), 2.35-2.18 (m, 1H), 2.18-1.89 (m, 1H), 0.90(s,9H)。

Embodiment 238

Synthesize (4S)-N- (6- methylpyrazine -2- bases) -7- (6- picoline -3- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4S) -7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysenes -1,4- that 30min is stirred at room temperature under a nitrogen Endo-methylene group pyrido [2,3-b] [1,4] diaza(600mg, 2.378mmol), triphosgene (706mg, 2.378mmol) and Added in solution of the triethylamine (1.989mL, 14.27mmol) in tetrahydrofuran (15mL) 6- methylpyrazine -2- amine (778mg, 7.13mmol).Reactant mixture is stirred into 16h at 70 DEG C, room temperature is subsequently cooled to.Reactant mixture is poured into water and is used in combination EtOAc extracts (3 × 15mL).Organic layer water, the aqueous salt solu-tion of merging, it is dried over sodium sulfate, and evaporated, obtain rough Compound (TLC eluant, eluents：5%MeOH/DCM:R_f-0.1；UV activation).Crude compound is purified through column chromatography, uses neutrality Aluminum oxide and with 30%EtOAc/ petroleum ethers elute, obtain pure (4S)-N- (6- methylpyrazine -2- bases) -7- (6- methyl pyrroles Pyridine -3- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (151mg, 0.389mmol, 16.38% yield), it is pale solid.LCMS(m/z):388.2[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.79 (s, 1H) 9.33 (s, 1H) 9.10 (d, J=2.41Hz, 1H) 8.55 (dd, J=8.11,2.41Hz, 1H) 8.19 (s, 1H) 7.62 (d, J=7.89Hz, 1H) 7.43 (d, J=7.89Hz, 1H) 7.31 (d, J=8.11Hz, 1H) 5.70 (dd, J=5.92,3.07Hz, 1H) 3.36-3.14 (m, 3H) 3.02 (dd, J= 11.95,3.40Hz,1H)2.65(s,3H)2.56(s,3H)2.42-2.32(m,1H)2.18-2.01(m,1H)。

Embodiment 239

(4S) -7- (6- cyclopropyl -5- (trifluoromethyl) pyridin-3-yl)-N- (pyrazine -2- bases) -3,4- dihydro -1 is synthesized, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines with argon-degassed 20min And [2,3-b] [1,4] diaza- 5 (2H)-formamides (600mg, 1.894mmol), 2- cyclopropyl -5- (4,4,5,5- tetramethyls Base -1,3,2- dioxaborolan -2- bases) -3- (trifluoromethyl) pyridine (mg, mmol) and K₃PO₄(804mg,3.79mmol) X-Phos (90mg, 0.189mmol), three (dibenzylidenes third are added in solution in the dioxane of Isosorbide-5-Nitrae-(20mL), water (3mL) Ketone) two palladiums (0) (87mg, 0.095mmol), argon-degassed 10min is used again.Reactant mixture is stirred into 16h at 100 DEG C, so After be cooled to room temperature.Reactant mixture is filtered through diatomite, then filtrate water dilutes and extracted (3 × 15mL) with EtOAc. Organic layer water, the aqueous salt solu-tion of merging, it is dried over sodium sulfate, and evaporated, obtain crude compound (TLC eluant, eluents：5% MeOH/DCM:R_f-0.2；UV activation).Crude compound is purified through column chromatography, using neutral alumina and uses 30%EtOAc/ Petroleum ether is eluted, and obtains pure (4S) -7- (6- cyclopropyl -5- (trifluoromethyl) pyridin-3-yl)-N- (pyrazine -2- bases) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (380mg, 0.813mmol, 42.85% yield), it is pale solid, LCMS (m/z):468.2[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.66 (s, 1H) 9.51 (d, J=1.53Hz, 1H) 9.08 (d, J= 1.97Hz, 1H) 8.58 (d, J=2.19Hz, 1H) 8.31-8.24 (m, 2H) 7.64 (d, J=7.89Hz, 1H) 7.38 (d, J= 7.89Hz, 1H) 5.71 (dd, J=5.92,3.07Hz, 1H) 3.34-3.14 (m, 3H) 3.03 (dd, J=12.06,3.29Hz, 1H)2.47-2.30(m,2H)2.15-2.04(m,1H)1.33-1.28(m,2H)1.16-1.10(m,2H)。

Embodiment 240

Synthesize (4S) -7- (2- cyclopropyl morpholino)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides

Under a nitrogen nitrine is added at 0 DEG C in THF (10mL) solution to pyrazine -2- formic acid (550mg, 4.43mmol) Simultaneously 2h is stirred at room temperature in diphenyl phosphate (2439mg, 8.86mmol) and TEA (3.09mL, 22.16mmol).Mixed to the reaction 2- cyclopropyl -4- ((4S) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas are added in thing- 7- yls) morpholine (888mg, 3.10mmol), then stirs 16h at 100 DEG C.Reactant mixture is diluted with water and ethyl acetate is used Extract (2x25ml).The organic layer aqueous salt solu-tion of merging, through anhydrous sodium sulfate drying and is concentrated under reduced pressure, and obtains roughization Compound.Through flash column chromatography, (100-200 silica gel, uses 2%MeOH/CH to crude product₂Cl₂Elution), obtain compound (4S)- 7- (2- cyclopropyl morpholino)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (90mg, 0.219mmol, 4.95% yield), it is Light brown solid.(TLC systems:5% methanol/ DCM.R_fValue：0.3), LCMS (m/z):408.33[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.32 (s, 1H), 9.37 (d, J=1.32Hz, 1H), 8.43-8.13 (m, 2H), 7.41 (d, J=8.55Hz, 1H), 6.56 (d, J=8.77Hz, 1H), 5.56-5.28 (m, 1H), 4.13-3.73 (m, 3H), 3.55 (td, J=11.56,2.52Hz, 1H), 3.16-2.63 (m, 7H), 2.31-1.99 (m, 1H), 1.85 (dt, J= 13.92,6.85Hz,1H),1.08-0.76(m,1H),0.59-0.12(m,4H)

Embodiment 241

Synthesize (4S) -7- (4,6- lutidines -3- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

By Cs₂CO₃(1237mg, 3.797mmol) and 2,4- dimethyl -5- (4,4,5,5- tetramethyl -1,3,2- dioxas The amyl- 2- yls of boron heterocycle) pyridine (590mg, 2.531mmol) adds to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (400mg, 1.265mmol) are in 1,4- dioxanes In solution in the mixture of (9mL) and water (1mL).By reactant mixture purification for argon 30min.By PdOAc₂(28.3mg, Above-mentioned reactant mixture 0.126mmol) is added to X-Phos (120.6mg, 0.253mmol), 16h are stirred at 100 DEG C.Will reaction Mixture is cooled to room temperature, is concentrated in vacuo and by residue distribution between water (40mL) and EtOAc (80mL).Separate organic layer And through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into crude compound.Crude compound is purified through column chromatography, is used Silica gel (100-200 mesh), 2% ethanol/methylene as eluant, eluent, obtain (4S) -7- (4,6- lutidines -3- bases) - N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (135mg, 0.332mmol, 52.5% yield), it is pale solid.(TLC eluant, eluents：10%MeOH/DCM Rf:0.4；UV Activation), LCMS (m/z):388.28[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.37 (s, 1H), 9.45 (d, J=1.32Hz, 1H), 8.54 (s, 1H), 8.22 (d, J=2.63Hz, 1H), 8.16-8.20 (m, 1H), 7.62 (d, J=7.67Hz, 1H), 7.12 (s, 1H), 7.07 (d, J =7.89Hz, 1H), 5.71 (dd, J=5.81,2.96Hz, 1H), 3.15-3.36 (m, 3H), 3.03 (dd, J=12.17, 3.40Hz,1H),2.59(s,3H),2.46(s,3H),2.28-2.38(m,1H),2.04-2.17(m,1H)。

Embodiment 242

Synthesize (4S)-N- (3,3- difluoros cyclobutyl) -7- (6- picoline -3- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Triphosgene (212mg, 0.714mmol) is slowly added to (4S) -7- (6- picolines -3- in batches in room temperature Base) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(300mg, 1.190mmol) is four In the solution of stirring in hydrogen furans (THF) (15mL).The reactant mixture is stirred into 30min.By DIPEA (0.623mL, Above-mentioned reactant mixture 3.57mmol) is added to 3,3- difluoro cyclobutyl amine (191mg, 1.785mmol), 16h are stirred at 80 DEG C. Reactant mixture is cooled to room temperature, is concentrated in vacuo and by residue distribution between water (40mL) and EtOAc (80mL).Separation Organic layer and through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into crude compound.Crude compound is pure through column chromatography Change, using silica gel (100-200 mesh), 2% ethanol/methylene obtains (4S)-N- (3,3- difluoro ring fourths as eluant, eluent Base) -7- (6- picoline -3- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (113mg, 0.283mmol, 28.5% yield), it is pale solid.(TLC eluant, eluents：10%MeOH/ DCM R_f:0.4；UV activation), LCMS (m/z):386.27[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 10.93 (br d, J=5.92Hz, 1H), 8.88 (d, J=2.19Hz, 1H), 7.94 (dd, J=8.11,2.41Hz, 1H), 7.56 (d, J=7.67Hz, 1H), 7.31-7.22 (m, 2H), 5.59 (dd, J =5.92,3.29Hz, 1H), 4.29 (br s, 1H), 3.00-3.29 (m, 5H), 2.95 (dd, J=11.95,3.40Hz, 1H), 2.66-2.50(m,5H),2.31-2.21(m,1H),2.06-1.98(m,1H)

Embodiment 243

Synthesis (4S) -7- (1- methyl -6- oxo -1,6- dihydropyridine -3- bases)-N- (pyrazine -2- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

28 DEG C by tripotassium phosphate (804mg, 3.79mmol) add to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (500mg, 1.579mmol) and 1- methyl- 5- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) pyridine -2 (1H) -one (445mg, 1.894mmol) exists In 1,4- dioxanes (10mL) and the solution of the stirring in water (2.000mL).Reactant mixture is deaerated 10min, then added Pd₂(dba)₃(72.3mg, 0.079mmol), and X-phos (75mg, 0.158mmol).Reactant mixture is deaerated 15min again. Reactant mixture is stirred 1 hour in microwave at 100 DEG C.Reactant mixture is cooled to 28 DEG C, then distributed in water Between (10mL) and ethyl acetate (2 × 15mL).Separating ethyl acetate layer, then through anhydrous Na₂SO₄Dry, filter and by filtrate Evaporation, obtains rough brown solid (TLC eluant, eluents：10%MeOH/EtOAc:R_f-0.2；UV activation).Thick material is through post color Spectrum purifying, use (100-200 mesh) silica gel simultaneously eluted with 2%MeOH/EtOAc, obtain (4S) -7- (1- methyl -6- oxo -1, 6- dihydropyridine -3- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (330mg, 0.840mmol, 53.2% yield), it is pale solid, LCMS (m/z):390.26[M +H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.78 (s, 1H), 9.61 (d, J=1.53Hz, 1H), 8.57 (d, J= 2.63Hz, 1H), 8.33 (d, J=2.63Hz, 1H), 8.22 (dd, J=2.52,1.64Hz, 1H), 7.88 (dd, J=9.54, 2.74Hz, 1H), 7.57 (d, J=7.89Hz, 1H), 7.18 (d, J=8.11Hz, 1H), 6.69 (d, J=9.43Hz, 1H), 5.69 (dd, J=5.92,3.07Hz, 1H), 3.78 (s, 3H), 3.12-3.30 (m, 3H), 3.01 (dd, J=12.06, 3.29Hz,1H),2.28-2.41(m,1H),2.00-2.13(m,1H)

Embodiment 244

Synthesis (4S) -7- (1- methyl -6- oxo -1,6- dihydropyridine -3- bases)-N- (pyridin-3-yl) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

28 DEG C by tripotassium phosphate (672mg, 3.17mmol) add to (4S) -7- chloro- N- (pyridin-3-yl) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (500mg, 1.583mmol), and 1- first Base -5- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) pyridine -2 (1H) -one (447mg, 1.900mmol) In the solution of stirring in 1,4- dioxanes (10mL) and water (2.000mL).Reactant mixture is deaerated 10min, Ran Houjia Enter Pd₂(dba)₃(72.5mg, 0.079mmol) and X-phos (75mg, 0.158mmol).Reactant mixture is deaerated 15min again. Reactant mixture is stirred 1 hour in microwave at 100 DEG C.Reactant mixture is cooled to 28 DEG C, then distributed in water Between (10mL) and EtOAc (25mL).EtOAc layers of separation, then through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain Thick material, it is brown solid (TLC eluant, eluents：10%MeOH/EtOAc:R_f-0.2；UV activation).Thick material is through column chromatography Purifying, using silica gel (100-200 mesh), product is eluted with 2%MeOH/ ethyl acetate, (4S) -7- (1- methyl -6- oxygen is obtained Generation -1,6- dihydropyridine -3- bases)-N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine- 5 (2H)-formamides (272mg, 0.679mmol, 42.9% yield), it is pale solid, LCMS (m/z): 389.27[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 12.93 (s, 1H), 8.61 (d, J=2.41Hz, 1H), 8.33 (dd, J= 4.71,1.43Hz, 1H), 8.05-8.20 (m, 1H), 7.74-7.87 (m, 2H), 7.57 (d, J=7.89Hz, 1H), 7.22- 7.33 (m, 1H), 7.09 (d, J=7.89Hz, 1H), 6.65-6.82 (m, 1H), 5.67 (dd, J=5.81,3.18Hz, 1H), 3.64 (s, 3H), 3.08-3.28 (m, 3H), 3.00 (dd, J=12.06,3.29Hz, 1H), 2.33 (qd, J=9.98, 4.49Hz,1H),2.00-2.12(m,1H)。

Embodiment 245

By Pd₂(dba)₃It is chloro- that (0.043g, 0.047mmol) and X-phos (0.036g, 0.095mmol) add to (4S) -7- N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (0.3g, 0.947mmol), the fluoro- 5- of 3- (4,4,5,5- tetramethyls -1,3,2- dioxaborolan -2- bases) pyridine -2- formonitrile HCNs (0.352g, 1.421mmol) and potassium dihydrogen phosphate (0.258g, 1.894mmol) are at 1,4- dioxanes (5mL):Taken off in water (1mL) In the solution of gas, 90 DEG C are then heated to, is kept for 15 hours.Room temperature is cooled to, is then filtered by Celite pad.Filtrate water (25mL) and ethyl acetate (50mL) dilute.The organic layer of separation is filtered and evaporated and obtain thick material through anhydrous sodium sulfate drying (TLC eluant, eluents：10%MeOH/ ethyl acetate；UV is activated；R_f:0.25).Crude compound is purified through column chromatography, uses neutrality Aluminum oxide and in 75% ethyl acetate/hexane elute, obtain (4S) -7- (6- cyano group -5- fluorine pyridin-3-yl)-N- (pyrazine - 2- yls) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1, -4] diaza- 5 (2H)-formamides (0.105g, 0.251mmol, 26.5% yield), it is pale solid, LCMS (m/z):403.18[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 13.57 (s, 1H), 9.53 (d, J=1.53Hz, 1H), 9.11 (t, J= 1.43Hz, 1H), 8.69 (dd, J=9.87,1.75Hz, 1H), 8.27-8.40 (m, 2H), 7.71 (d, J=8.11Hz, 1H), 7.54 (d, J=8.11Hz, 1H), 5.71 (dd, J=5.81,3.18Hz, 1H), 3.01-3.37 (m, 4H), 2.32-2.45 (m, 1H),2.01-2.21(m,1H)。

Embodiment 246

Synthesize (4S) -7- (6- cyanopyridine -3- bases)-N- (pyridin-3-yl) -3,4- dihydros -1,4- methylene-pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4S) -7- chloro- N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene group pyridines with argon-degassed 20min And [2,3-b] [1,4] diaza- 5 (2H)-formamides (600mg, 1.900mmol), 5- (4,4,5,5- tetramethyls -1,3,2- Dioxaborolan -2- bases) pyridine -2- formonitrile HCNs (525mg, 2.280mmol) and K₃PO₄(807mg, 3.80mmol) is in 1,4- X-phos (91mg, 0.190mmol), three (dibenzalacetone) two palladium are added in solution in dioxane (20mL), water (3mL) (0) (87mg, 0.095mmol), uses argon-degassed 10min again.Reactant mixture is stirred 16 hours at 100 DEG C, Ran Houleng But to room temperature.Reactant mixture is filtered through diatomite, then filtrate water dilutes and extracted (3 × 20mL) with EtOAc.Merge Organic layer water, aqueous salt solu-tion, it is dried over sodium sulfate, and evaporated, obtain crude compound (TLC eluant, eluents：5% MeOH/DCM:R_f-0.3；UV activation).Crude compound is purified through column chromatography, using neutral alumina and uses 30-40% EtOAc/ Hex, obtains pure (4S) -7- (6- cyanopyridine -3- bases)-N- (pyridin-3-yl) -3,4- dihydros-Isosorbide-5-Nitrae-bridge Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (399mg, 1.033mmol, 54.4% yield), its For pale solid, LCMS (m/z):384.3[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 12.71 (s, 1H), 9.17 (dd, J=2.2,0.7Hz, 1H), 8.54 (d, J= 2.4Hz, 1H), 8.33 (dd, J=4.7,1.4Hz, 1H), 8.23 (dd, J=8.0,2.3Hz, 1H), 8.07-8.12 (m, 1H), 7.87 (d, J=8.1Hz, 1H), 7.69 (d, J=8.1Hz, 1H), 7.38 (d, J=7.9Hz, 1H), 7.26-7.31 (m, 1H), 5.71 (dd, J=5.9,2.8Hz, 1H), 3.12-3.34 (m, 3H), 3.01-3.09 (m, 1H), 2.36 (m, 1H), 2.1 (m, 1H)。

Embodiment 247

Synthesize (4S) -7- ((S) -3- methyl morpholine generations)-N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

By Cs₂CO₃(3.10g, 9.50mmol), (S) -3- methyl morpholines (0.641g, 6.33mmol) are added in seal pipe (4S) -7- chloro- N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 In solution of (the 2H)-formamide (1.0g, 3.17mmol) in the dioxane of Isosorbide-5-Nitrae-(10mL), while using purification for argon 25min.With Afterwards by PdOAc₂(0.142g, 0.633mmol) and X-phos (0.604g, 1.267mmol) add to reactant mixture, Ran Hou 100 DEG C of stirring 4h.Reactant mixture is cooled to room temperature, removal of solvent under reduced pressure, by residue distribution in water (40mL) and EtOAc Between (80mL).Separate organic layer and through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain crude compound.Used Column chromatography is purified, and using silica gel (100-200 mesh), 2% ethanol/methylene obtains (4S) -7- ((S) -3- as eluant, eluent Methyl morpholine generation)-N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (140mg, 0.366mmol, 11.56% yield), it is pale solid.(TLC eluant, eluents：10%MeOH/ DCM,R_f:0.4；UV activation), LCMS (m/z):381.30[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 12.42 (s, 1H), 8.50 (d, J=2.41Hz, 1H), 8.31 (d, J= 4.60Hz, 1H), 8.16 (br d, J=8.55Hz, 1H), 7.39 (d, J=8.55Hz, 1H), 7.31-7.26 (m, 1H), 6.24 (d, J=8.55Hz, 1H), 5.60 (dd, J=5.92,3.29Hz, 1H), 4.06 (brdd, J=11.18,3.51Hz, 2H), 3.85-3.79 (m, 2H), 3.69-3.51 (m, 2H), 3.37-3.04 (m, 4H), 2.91 (dd, J=11.84,3.29Hz, 1H), 2.31-2.18 (m, 1H), 2.07-1.97 (m, 1H), 1.27 (d, J=6.80Hz, 3H)

Embodiment 248

Synthesize (4S)-N- (4- picoline -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaThree are added in the solution of the stirring of (400mg, 1.585mmol) in THF (15mL, in seal pipe) Phosgene (282mg, 0.951mmol), and stirring 30min, then add triethylamine (1.105mL, 7.93mmol) and 4- methyl pyrroles Pyridine -2- amine (257mg, 2.378mmol) simultaneously heats 15h at 65 DEG C.Reactant mixture is cooled to room temperature.THF is distilled out, then Distribution is between water (25mL) and EtOAc (40mL).Separate organic layer and through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, Obtain crude compound.Crude compound is through flash column chromatography (silica gel:100-200 mesh, eluant, eluent：2% methanol/DCM), 600mg is obtained, LCMS purity is 60%.Crude compound purifies (condition through preparation HPLC：MP-A:10mM ammonium hydrogen carbonate (water Solution) MP-B：Acetonitrile post：The μ methods of X-Bridge (250x30) mm 10：0/30,10/50,10.1/30,15/30 flow velocity： 30ml/min eluant, eluents：THF+ACN+MeOH), (4S)-N- (4- picoline -2- bases) -7- (2- picolines -4- are obtained Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (210mg, 0.541mmol, 34.1% yield), it is pale solid (TLC eluant, eluents：10%MeOH/EtOAc, R_f:0.4), LCMS (m/ z):387.27[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δppm:13.48 (s, 1H), 8.61 (d, J=5.26Hz, 1H), 8.25-8.17 (m, 2H), 8.07 (s, 1H), 7.71 (d, J=3.95Hz, 1H), 7.61 (d, J=7.89Hz, 1H), 7.48 (d, J=8.11Hz, 1H), 6.85 (d, J=4.82Hz, 1H), 5.70 (dd, J=5.70,3.51Hz, 1H), 3.33-3.14 (m, 3H), 3.14-2.89 (m,1H),2.74(s,3H),2.41-2.32(m,4H),2.13-2.00(m,1H)。

Embodiment 249

Synthesize (4S)-N- (6- isopropylpyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

In the solution of stirring of the room temperature to 6- isopropylpyrazine -2- amine (141mg, 1.030mmol) in THF (5mL) Add triethylamine (0.663mL, 4.76mmol) and triphosgene (235mg, 0.793mmol) and stirring 30min.Then add (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza (200mg, 0.793mmol), then heats 16h at 80 DEG C.Reactant mixture is cooled to room temperature, then distributed at water (25mL) Between EtOAc (40mL).Separate organic layer and through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain rough chemical combination Thing.Crude mixture is through flash column chromatography (silica gel:100-200 mesh, eluant, eluent：2% methanol/DCM), obtain (4S)-N- (6- isopropylpyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (60mg, 0.144mmol, 18.15% yield), it is pale solid (TLC:Elution Agent：5% methanol/DCM, R_f:0.4), LCMS (m/z):416.3[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.34 (s, 1H), 9.37 (s, 1H), 8.70 (d, J=5.26Hz, 1H), 8.21 (s, 1H), 8.00 (br d, J=3.73Hz, 1H), 7.60-7.69 (m, 2H), 7.47 (d, J=8.11Hz, 1H), 5.73 (dd, J=5.81,3.18Hz, 1H), 3.14-3.37 (m, 3H), 2.98-3.09 (m, 2H), 2.66 (s, 3H), 2.27-2.46 (m, 1H), 2.09 (dt, J=14.03,7.23Hz, 1H), 1.33 (dd, J=7.02,1.10Hz, 6H).

Embodiment 250

Synthesize (4S)-N- (pyridine -2- bases) -7- (3- (trifluoromethyl) pyrrolidin-1-yl) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -7- (3- (trifluoromethyl) pyrrolidin-1-yl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza(peak 1, the separation from intermediate SFC) (440mg, 1.475mmol) is in tetrahydrofuran (THF) NaH (295mg, 7.37mmol) is added in the solution in (10mL).3- (pyridine -2- bases) -2H- pyridines are added after 30min And [1,2-a] [1,3,5] triazine -2,4 (3H)-diketone (531mg, 2.212mmol) and by reactant mixture 70 DEG C stirring 16h.Reactant mixture is poured on cold water (50mL) and is extracted with ethyl acetate (2x30mL).Organic layer is dry through anhydrous sodium sulfate It is dry and be concentrated under reduced pressure, obtain crude product.Crude product is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：90%Hex/ EtOAc), (4S)-N- (pyridine -2- bases) -7- (3- (trifluoromethyl) pyrrolidin-1-yl) -3,4- dihydro-Isosorbide-5-Nitrae-bridge methylenes are obtained Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (266.8mg, 0.631mmol, 42.8% yield), it is Pale solid.(TLC systems:R_f:04,:EtOAc), LCMS (m/z):419.2[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.33 (s, 1H), 8.19 (d, J=3.73Hz, 1H), 8.15 (d, J= 8.33Hz, 1H), 7.66-7.60 (m, 1H), 7.32 (d, J=8.55Hz, 1H), 6.91 (dd, J=6.58,5.04Hz, 1H), 5.96 (d, J=8.33Hz, 1H), 5.58 (dd, J=6.03,3.18Hz, 1H), 4.01-3.95 (m, 1H), 3.86 (dd, J= 11.29,7.34Hz, 1H), 3.66 (td, J=8.99,4.60Hz, 1H), 3.60-3.53 (m, 1H), 3.23-3.05 (m, 4H), 2.89 (dd, J=11.84,3.51Hz, 1H), 2.37-2.18 (m, 3H), 1.98 (dt, J=13.81,6.91Hz, 1H).

Embodiment 251

Synthesis (4S) -7- (1- methyl -3- (trifluoromethyl) -1H- pyrazoles -5- bases)-N- (pyrazine -2- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

By the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (0.400g, 1.263mmol), (4,4,5,5- tetramethyls -1,3,2- dioxa boron is miscellaneous by 1- methyl -5- The amyl- 2- yls of ring) -3- (trifluoromethyl) -1H- pyrazoles (0.697g, 2.53mmol) and tripotassium phosphate (0.803g, 3.79mmol) exist Then suspension argon gas in the dioxane of Isosorbide-5-Nitrae-(25mL) and water (4mL) adds X-phos in room temperature degassing 15min (0.0601g, 0.126mmol) and Pd (dba)₂(0.046g,0.051mmol).Reactant mixture is deaerated again 15min and 100 DEG C are stirred 16 hours.(indicated after the completion of reaction by TLC), reactant mixture is cooled to 28 DEG C and is diluted with water (15mL) and extracted (2X25mL) with EtOAc.Organic layer priority water and aqueous salt solu-tion, it is then dry through anhydrous sodium sulfate It is dry, filter and filtrate evaporation is obtained into thick material (TLC eluant, eluents：100% ethyl acetate/hexane, R_fValue：0.3, UV activation ).Thick material is purified through column chromatography, using neutral alumina, with 40% ethyl acetate/petroleum ether eluted product, obtain (4S)- 7- (1- methyl -3- (trifluoromethyl) -1H- pyrazoles -5- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides (0.208g, 0.482mmol, 38.2% yield), it is solid for white Body, LCMS (m/z):431.23[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 13.08 (s, 1H), 9.51 (d, J=1.5Hz, 1H), 8.29 (d, J=2.6Hz, 1H), 8.25-8.22 (m, 1H), 7.65 (d, J=7.9Hz, 1H), 7.21 (d, J=7.9Hz, 1H), 7.05 (s, 1H), 5.71 (dd, J=5.8,3.2Hz, 1H), 4.24 (s, 3H), 3.33-3.15 (m, 3H), 3.04 (dd, J=12.2,3.2Hz, 1H), 2.42-2.30 (m, 1H), 2.10 (dt, J=14.1,6.9Hz, 1H).

Embodiment 252

Synthesize (4S) -7- cyano group-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] Diaza- 5 (2H)-formamides

By X-phos (0.018g, 0.047mmol) and ((2,6- the diisopropyl phenyls) -4,5- glyoxalidine of 1,3- bis- -2- Subunit) chlorine) (3- phenyl allyls) palladium (2) (0.031g, 0.047mmol) adds to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3b] [1,4] diaza- 5 (2H)-formamides (0.3g, 0.947mmol) and iron cyanogen Change potassium (0.174g, 0.474mmol) at 1,4- dioxanes (5mL):In water (1mL) in the solution of degassing, 100 are then heated to DEG C, keep 15h.Room temperature is cooled to, is then filtered through Celite pad.Filtrate water (25mL) and ethyl acetate (100mL) dilution. The organic layer of separation is filtered and evaporated through anhydrous sodium sulfate drying, obtains crude compound (TLC eluant, eluents：100% acetic acid second Ester；UV is activated；R_f~0.3).Crude mixture is purified through column chromatography using aluminum oxide and washed in 50% ethyl acetate/hexane It is de-, obtain (4S) -7- cyano group-N- (pyrazine -2- bases) -3,4- dihydros-Isosorbide-5-Nitrae-endo-methylene group pyrido [2,3-b] [Isosorbide-5-Nitrae] phenodiazine It is miscellaneous- 5 (2H)-formamides (0.13g, 0.415mmol, 43.8% yield), it is white solid, LCMS (m/z):308.19[M +H]⁺。

¹H NMR(400MHz,CDCl₃):δppm 12.48(br s,1H),9.45(s,1H),8.32(s,2H),7.62(d, J=7.89Hz, 1H), 7.37 (d, J=7.89Hz, 1H), 5.68 (dd, J=5.81,3.18Hz, 1H), 3.24 (t, J= 7.56Hz, 2H), 2.99-3.14 (m, 2H), 2.36 (td, J=14.09,6.03Hz, 1H), 2.01-2.16 (m, 1H).

Embodiment 253

Synthesize (4S) -7- (1- isopropyl -1H- pyrazoles -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines with argon-degassed 20min And [2,3-b] [1,4] diaza- 5 (2H)-formamides (500mg, 1.579mmol), 1- isopropyls -4- (4,4,5,5- tetramethyls Base -1,3,2- dioxaborolan -2- bases) -1H- pyrazoles (447mg, 1.894mmol) and K₃PO₄(670mg,3.16mmol) X-phos (75mg, 0.158mmol), three (dibenzylidenes third are added in solution in the dioxane of Isosorbide-5-Nitrae-(15mL), water (3mL) Ketone) two palladiums (0) (72.3mg, 0.079mmol), argon-degassed 5min is used again.Reactant mixture is small in 100 DEG C of stirrings 16 When, it is subsequently cooled to room temperature.Reactant mixture is filtered through diatomite, then filtrate water dilute and extracted with EtOAc (3 × 20mL).Organic layer water, the aqueous salt solu-tion of merging, dried over sodium sulfate, and evaporated, obtaining crude compound, (TLC is eluted Agent：5%MeOH/DCM:R_f-0.2；UV activation).Crude compound is purified through column chromatography, using neutral alumina and uses 30- 40%EtOAc/ Hex, obtains pure (4S) -7- (1- isopropyl -1H- pyrazoles -4- bases)-N- (pyrazine -2- bases) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (418mg, 1.070mmol, 67.8% yield), it is pale solid, LCMS (m/z):391.2[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 14.14 (s, 1H), 9.62 (d, J=1.5Hz, 1H), 8.64 (s, 1H), 8.26- 8.32 (m, 2H), 8.02 (s, 1H), 7.50 (d, J=7.9Hz, 1H), 7.16 (d, J=8.1Hz, 1H), 5.67 (dd, J=5.8, 3.2Hz, 1H), 4.61 (m, 1H), 3.12-3.31 (m, 3H), 2.99 (dd, J=11.9,3.2Hz, 1H), 2.32 (m, 1H), 2.06 (m, 1H), 1.63 (d, J=6.8Hz, 6H).

Embodiment 254

Synthesize (4S) -7- (6- picoline -3- bases)-N- (tetrahydrofuran -3- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaTriphosgene is added in the solution of (300mg, 1.189mmol) in tetrahydrofuran (THF) (10mL) (176mg,0.594mmol.Addition DIPEA (1.038mL, 5.94mmol) after 30min, tetrahydrofuran -3- amine (155mg, 1.783mmol) and by reactant mixture at 70 DEG C stir 16h.So that reactant mixture reaches room temperature, cold water is then poured on On (50mL) and it is extracted with ethyl acetate (2x30mL).Organic layer is concentrated under reduced pressure through anhydrous sodium sulfate drying, obtains crude product. Crude product is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：3%MeOH/DCM), (4S) -7- (6- methyl is obtained Pyridin-3-yl)-N- (tetrahydrofuran -3- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (137.1mg, 0.371mmol, 31.2% yield), it is pale solid.(TLC systems:R_f:0.2,5% MeOH-DCM), LCMS (m/z):366.2[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 10.76 (br t, J=6.80Hz, 1H), 8.89 (s, 1H), 7.99 (dd, J=8.22,2.08Hz, 1H), 7.54 (d, J=7.89Hz, 1H), 7.30-7.21 (m, 2H), 5.62 (dd, J=5.81, 2.96Hz, 1H), 4.56 (br dd, J=4.82,1.97Hz, 1H), 4.01-3.75 (m, 4H), 3.30-3.06 (m, 3H), 2.99-2.91(m,1H),2.62(s,3H),2.39-2.21(m,2H),2.07-1.85(m,2H)。

Embodiment 255

Synthesize (4S) -7- (the fluoro- 6- picolines -3- bases of 5-)-N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Tripotassium phosphate (323mg, 1.520mmol) is added into (4S) -7- chloro- N- (pyridin-3-yl) -3,4- two at 25 DEG C Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (240.0mg, 0.760mmol) and 3- Fluoro- 2- methyl -5- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) pyridine (234mg, 0.988mmol) exists In solution in 1,4- dioxanes (5.0mL) and water (1.0mL).Reactant mixture is deaerated 10min.By X-Phos (36.2mg, 0.076mmol) and Pd₂(dba)₃(34.8mg, 0.038mmol) adds to reactant mixture and the 10min that deaerates again.By reaction mixing Thing is stirred 1 hour at 100 DEG C in microwave.Reactant mixture is filtered by Celite pad and washed with ethyl acetate.Filtrate With water and aqueous salt solu-tion.The organic phase of separation through anhydrous sodium sulfate drying, filter and evaporate obtain thick material (TLC elute Agent：5%MeOH/DCM；UV is activated；R_f~0.3).The purification of crude product on GRACE posts, is eluted with 3-5%MeOH/DCM, is obtained Pure (4S) -7- (the fluoro- 6- picolines -3- bases of 5-)-N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (122.0mg, 0.312mmol, 41.0% yield), it is solid for canescence Body, LCMS (m/z):391.30[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 12.91 (s, 1H), 8.77 (t, J=1.42Hz, 1H), 8.62 (d, J= 2.41Hz, 1H), 8.32 (dd, J=4.71,1.43Hz, 1H), 8.12 (ddd, J=8.33,2.63,1.53Hz, 1H), 7.73 (dd, J=9.87,1.75Hz, 1H), 7.64 (d, J=7.89Hz, 1H), 7.27-7.32 (m, 2H), 5.70 (dd, J=5.92, 3.29Hz, 1H), 3.14-3.34 (m, 3H), 3.03 (dd, J=12.06,3.29Hz, 1H), 2.62 (d, J=3.07Hz, 3H), 2.28-2.38(m,1H),2.05-2.14(m,1H)

Embodiment 256

Synthesize (4S) -7- (the fluoro- 6- picolines -3- bases of 5-)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides:

Tripotassium phosphate (402mg, 1.894mmol) is added into the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- two at 25 DEG C Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (300.0mg, 0.947mmol), 3- Fluoro- 2- methyl -5- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) pyridine (292mg, 1.231mmol) exists In solution in 1,4- dioxanes (5.0mL) and water (1.0mL).Reactant mixture is deaerated 10min.By X-Phos (45.2mg, 0.095mmol) and Pd₂(dba)₃(43.4mg, 0.047mmol) adds to reactant mixture, and then deaerate 10min again.Reaction is mixed Compound is stirred 1 hour at 100 DEG C in microwave.Reactant mixture is filtered by Celite pad and washed with ethyl acetate.Filter Liquid water and aqueous salt solu-tion, the organic phase of separation are filtered and evaporated and obtain thick material (TLC is washed through anhydrous sodium sulfate drying De- agent：5%MeOH/DCM；UV is activated；R_f~0.3).Purification of crude product exists on GRACE posts, uses 3-5%MeOH/ dichloromethane Elution, obtains pure (4S) -7- (the fluoro- 6- picolines -3- bases of 5-)-N- (pyrazine -2- bases) -3,4- dihydro-Isosorbide-5-Nitrae-bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (150.0mg, 0.365mmol, 38.5% yield), it is Pale solid, LCMS (m/z):392.3[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 13.79 (s, 1H), 9.53 (d, J=1.5Hz, 1H), 8.88 (t, J=1.5Hz, 1H), 8.38 (dd, J=10.7,1.97Hz, 1H), 8.30-8.34 (m, 2H), 7.64 (d, J=7.9Hz, 1H), 7.44 (d, J= 8.1Hz, 1H), 5.70 (dd, J=6.03,3.2Hz, 1H), 3.1-3.3 (m, 3H), 3.03 (dd, J=12.1,3.3Hz, 1H), 2.62 (s, 3H), 2.36 (dddd, J=14.1,9.9,5.9,4.1Hz, 1H), 2.04-2.15 (m, 1H)

Embodiment 257

Synthesize (4S) -7- (1- cyclopropyl -1H- pyrazoles -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

By Pd₂(dba)₃It is chloro- that (0.087g, 0.095mmol) and X-phos (0.018g, 0.047mmol) add to (4S) -7- N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (0.3g, 0.947mmol), 1- cyclopropyl -4- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) -1H- pyrroles Azoles (0.333g, 1.421mmol) and potassium dihydrogen phosphate (0.258g, 1.894mmol) are at 1,4- dioxanes (5mL):In water (1mL) In the solution of degassing, 90 DEG C are then heated to, is kept for 15 hours.It is cooled to room temperature and is filtered by Celite pad.Filtrate water (25mL) and ethyl acetate (50mL) dilute.The organic layer of separation is filtered and evaporated and obtain thick material through anhydrous sodium sulfate drying (TLC eluant, eluents：10%MeOH/ ethyl acetate；UV is activated；R_f~0.30).Crude compound is purified through column chromatography, uses neutrality Aluminum oxide and in 75% ethyl acetate/hexane elute, obtain (4S) -7- (1- cyclopropyl -1H- pyrazoles -4- bases)-N- (pyrazine - 2- yls) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3b] [1,4] diaza- 5 (2H)-formamides (0.13g, 0.332mmol, 35.0% yield) pale solid, LCMS (m/z):389.32[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 14.14 (s, 1H), 9.60 (d, J=1.10Hz, 1H), 8.61 (s, 1H), 8.29-8.33 (m, 2H), 8.03 (s, 1H), 7.47-7.58 (m, 1H), 7.13 (d, J=8.11Hz, 1H), 5.66 (dd, J= 6.03,3.18Hz, 1H), 3.73 (tt, J=7.43,3.75Hz, 1H), 3.08-3.34 (m, 3H), 2.99 (dd, J=11.95, 3.18Hz, 1H), 2.32 (dddd, J=14.11,10.00,6.03,4.06Hz, 1H), 2.02-2.11 (m, 1H), 1.22-1.32 (m,2H),1.07-1.18(m,2H).

Embodiment 258

Synthesize (4S) -7- ((2S, 6R) -2,6- thebaine generations)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Diisopropylethylamine (3.34mL, 19.14mmol) is added into pyrazine -2- formic acid under argon gas stirring in room temperature In the solution of the stirring of (0.475g, 3.83mmol) in tetrahydrofuran (30mL).By diphenyl phosphate azide (1.053g, 3.83mmol) add to reactant mixture.2hr is stirred at room temperature in reactant mixture, then, by (2S, 6R) -2,6- dimethyl - 4- ((4S) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diazas- 7- bases) morpholine (0.735g, Reactant mixture 2.68mmol) is added to, is then stirred 16 hours at 65 DEG C.Reactant mixture is cooled to room temperature, then distributed Between water (20mL) and EtOAc (70mL).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, Thick material is obtained, it is brown solid (TLC eluant, eluents：100%EtOAc:R_f-0.3；UV activation).Crude residue is through post Chromatogram purification, using neutral alumina and uses 15%EtOAc/ Hex, obtains pure (4S) -7- ((2S, 6R) -2,6- bis- Methyl morpholine generation)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (0.132g, 0.329mmol, 8.58% yield), it is pale solid, LCMS (m/z):396.3[M+H ]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.39 (s, 1H), 9.56 (d, J=1.10Hz, 1H), 8.26 (s, 1H), 8.25 (s, 1H), 7.39 (s, 1H), 6.30 (s, 1H), 5.63 (dd, J=5.92,3.29Hz, 1H), 4.01 (br d, J= 12.50Hz, 2H), 3.68-3.89 (m, 2H), 3.16-3.28 (m, 1H), 3.05-3.15 (m, 2H), 2.92 (dd, J=11.84, 3.29Hz, 1H), 2.61 (t, J=11.62Hz, 2H), 2.27 (dddd, J=13.89,9.95,6.08,3.62Hz, 1H), 1.93-2.06 (m, 1H), 1.33 (d, J=6.14Hz, 6H)

Embodiment 259

Synthesize (4S)-N- (pyrazine -2- bases) -7- (2- (trifluoromethyl) pyrimidine-4-yl) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (300mg, 0.947mmol), 4- (4,4,5,5- tetramethyl -1,3,2- dioxaborolans -2- Base) -2- (trifluoromethyl) pyrimidine (260mg, 0.947mmol) and K₃PO₄(603mg, 2.84mmol) is at 1,4- dioxanes (20mL) In solution in add water (3mL).By reactant mixture argon gas in room temperature degassing 15min.Then X-Phos is added thereto (45.2mg, 0.095mmol) and Pd₂(dba)₃(43.4mg,0.047mmol).Reactant mixture is stirred into 16hr at 90 DEG C.Instead Process is answered to be monitored through TLC.Reactive material is filtered and is concentrated under reduced pressure through diatomite.Extract by residue diluted with water and with ethyl acetate Take (2 × 50mL).The organic layer priority water and aqueous salt solu-tion of merging simultaneously use sodium sulphate drying and dehydrating, filter and concentrate. Crude compound purifies (neutral alumina) product through column chromatography and eluted with 30% ethyl acetate/hexane.The fraction of collection is subtracted Pressure evaporation, then in high vacuum dry, obtains (4S)-N- (pyrazine -2- bases) -7- (2- (trifluoromethyl) pyrimidine-4-yl) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (230mg, 0.534mmol, 56.4% yield), it is pale solid, LCMS (m/z):429.3[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.60 (s, 1H), 9.59 (d, J=1.32Hz, 1H), 9.08 (d, J= 5.26Hz, 1H), 8.83 (d, J=5.26Hz, 1H), 8.22-8.47 (m, 3H), 7.75 (d, J=7.89Hz, 1H), 5.72 (dd, J=5.92,3.29Hz, 1H), 3.15-3.37 (m, 3H), 3.00-3.10 (m, 1H), 2.28-2.46 (m, 1H), 2.03-2.18 (m,1H)。

Embodiment 260

Synthesize (4S) -7- (1- methyl isophthalic acid H- pyrazole-3-yls)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

By the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous(4,4,5,5- tetramethyl -1,3,2- dioxa boron is miscellaneous by -5 (2H)-formamides (0.350g, 1.105mmol), 1- methyl -3- The amyl- 2- yls of ring) -1H- pyrazoles (0.690g, 3.31mmol) and tripotassium phosphate (0.703g, 3.31mmol) be in 1,4- dioxanes Deaerated 15 minutes in room temperature with argon gas in suspension in (25mL) and water (4mL).Then, X- is added into said mixture Phos (53mg, 0.110mmol) and Pd (dba)₂(0.032g, 0.055mmol) and deaerate again 15 minutes.Reactant mixture is existed 100 DEG C are stirred 16 hours.(indicated after the completion of reaction by TLC), be cooled to 28 DEG C and (15mL) is diluted with water and is extracted with EtOAc Take (2x25mL).Organic layer priority water and aqueous salt solu-tion, then through anhydrous sodium sulfate drying, filter and evaporate filtrate Obtain crude residue (TLC eluant, eluents：5%MeOH/DCM, R_fValue：0.3, UV activation).Crude residue is pure through column chromatography Change, using neutral alumina, with 50% ethyl acetate/petroleum ether eluted product, obtain (4S) -7- (1- methyl isophthalic acid H- pyrazoles -3- Base)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine (0.192g, 0.527mmol, 47.7% yield), it is pale solid, LCMS (m/z):363.30[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.98 (s, 1H), 9.56 (d, J=1.32Hz, 1H), 8.33-8.27 (m, 2H), 7.66 (d, J=8.11Hz, 1H), 7.56-7.52 (m, 1H), 7.46 (d, J=2.19Hz, 1H), 7.42 (d, J= 2.41Hz, 1H), 5.68 (dd, J=5.92,3.29Hz, 1H), 3.99 (s, 3H), 3.33-3.12 (m, 3H), 2.99 (dd, J= 12.06,3.29Hz,1H),2.36-2.26(m,1H),2.11-1.98(m,1H)

Embodiment 261

Synthesize (4S)-N- (5- ethyoxyl pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaAdded in the solution of the stirring of (100mg, 0.396mmol) in THF (10mL, in seal pipe) DMAP (145mg, 1.189mmol) and (5- ethyoxyl pyrazine -2- bases) phenyl carbamates (308mg, 1.189mmol), so Afterwards 16h is heated at 90 DEG C.Reactant mixture is cooled to room temperature, then distributed between water (25mL) and EtOAc (40mL).Point From organic layer and through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain crude compound.Crude residue is through quick post Chromatogram purification (silica gel:100-200 mesh, eluant, eluent：2% methanol/DCM), obtain (4S)-N- (5- ethyoxyl pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine (38mg, 0.090mmol, 22.79% yield), it is pale solid (TLC eluant, eluents：5% methanol/DCM R_f:0.4), LCMS(m/z):418.36[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.50 (s, 1H), 9.01 (d, J=1.32Hz, 1H), 8.61 (d, J= 5.26Hz, 1H), 8.06 (s, 1H), 7.98 (m, 1H), 7.63 (m, 2H), 7.47 (d, J=7.89Hz, 1H), 5.71 (dd, J= 5.92,3.07Hz, 1H), 4.40 (q, J=7.02Hz, 2H), 3.34-3.10 (m, 3H), 3.01 (dd, J=5.94,2.96Hz, 1H), 2.67 (s, 3H), 2.36-2.10 (m, 2H), 1.42 (t, J=7.13Hz, 3H).

Embodiment 262

Synthesize (4S) -7- ((2S, 6S) -2,6- thebaine generations)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Diisopropylethylamine (3.52mL, 20.15mmol) is added into pyrazine -2- formic acid (0.500g, 4.03mmol) four In the solution of stirring in hydrogen furans (30mL), and it is stirred at room temperature under argon gas.By diphenyl phosphate azide (1.109g, Reactant mixture 4.03mmol) is added to, is then stirred at room temperature 2 hours.Then, by (2S, 6S) -2,6- dimethyl -4- ((4S) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 7- bases) morpholine (0.774g, Reactant mixture 2.82mmol) is added to, is then stirred 16 hours at 65 DEG C.Reactant mixture is cooled to room temperature, then distributed Between water (20mL) and EtOAc (70mL).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, Crude residue is obtained, it is brown solid (TLC eluant, eluents：100%EtOAc:R_f-0.3；UV activation).Thick material is through post Chromatogram purification, using neutral alumina and uses 20%EtOAc/ Hex, obtains pure (4S) -7- ((2S, 6S) -2,6- bis- Methyl morpholine generation)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (0.321g, 0.799mmol, 19.82% yield), it is white solid, LCMS (m/z):396.3[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.41 (s, 1H), 9.56 (d, J=1.53Hz, 1H), 8.25 (d, J= 2.41Hz, 1H), 8.16 (dd, J=2.41,1.53Hz, 1H), 7.37 (d, J=8.55Hz, 1H), 6.25 (d, J=8.55Hz, 1H), 5.63 (dd, J=6.03,3.18Hz, 1H), 4.11-4.31 (m, 2H), 3.67 (dd, J=12.50,3.29Hz, 2H), 3.19-3.34 (m, 3H), 3.06-3.18 (m, 2H), 2.92 (dd, J=11.84,3.29Hz, 1H), 2.27 (dddd, J= 13.95,9.95,6.03,3.84Hz, 1H), 1.92-2.07 (m, 1H), 1.34 (d, J=6.36Hz, 6H)

Embodiment 263

Synthesize (4S) -7- (2- picoline -4- bases)-N- (pyridazine -4- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature to the chloro- N- of (4S) -7- (pyridazine -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diazaThe solution of -5 (2H)-formamides (0.3g, 0.947mmol) degassing in 1,4- dioxanes (10ml)/water (2ml) Middle addition (2- picoline -4- bases) boric acid (0.195g, 1.421mmol) and K₃PO₄(0.402g, 1.894mmol), three (two Asias Benzylacetone) (0.090g, 0.189mmol heat 16h at 100 DEG C by two palladiums (0) (0.087g, 0.095mmol) and x-phos.Will Reactant mixture is cooled to room temperature and is extracted with ethyl acetate (2x50mL).The organic layer of merging is through anhydrous Na₂SO₄Dry, filtering And evaporate filtrate, obtain crude compound.Crude compound is through flash column chromatography (silica gel:100-200 mesh, eluant, eluent： 3% methanol/EtOAc), obtain (4S) -7- (2- picoline -4- bases)-N- (pyridazine -4- bases) -3,4- dihydro-Isosorbide-5-Nitrae-bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (125mg, 0.333mmol, 35.1% yield), it is ash White solid (TLC eluant, eluents：10% methanol/EtOAc, R_f:0.4), LCMS (m/z):374.21[M+H]⁺。

¹H NMR(400MHz,CDCl3):δppm:13.50 (s, 1H), 9.10 (d, J=1.97Hz, 1H), 8.93-9.05 (m, 1H), 8.72 (d, J=5.04Hz, 1H), 8.00 (dd, J=5.92,2.85Hz, 1H), 7.69 (d, J=8.11Hz, 1H), 7.61-7.53 (m, 1H), 7.48 (dd, J=5.26,1.32Hz, 1H), 7.41 (d, J=7.58Hz, 1H), 5.67 (dd, J= 5.92,3.07Hz, 1H), 3.35-3.13 (m, 3H), 3.05 (dd, J=12.17,3.18Hz, 1H), 2.37 (dddd, J= 14.17,9.84,5.81,4.17Hz,3H),2.39(m,1H),2.15(m,1H)。

Embodiment 264

Synthesize (4S) -7- (2- picoline -4- bases)-N- (4- methylpyrimidine -2- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diazaTriphosgene is added in the solution of the stirring of (400mg, 1.585mmol) in THF (15mL, seal pipe) (282mg, 0.951mmol) is in room temperature and stirring 30min.Then add TEA (1.105mL, 7.93mmol) and 4- methylpyrimidines- 2- amine (260mg, 2.378mmol), 15h is heated at 65 DEG C.Reactant mixture is cooled to room temperature；THF is distilled out, Ran Houfen Fit between water (25mL) and DCM (40mL).Separate organic layer and through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain Crude compound.Crude compound is through flash column chromatography (silica gel:100-200 mesh, eluant, eluent：2% methanol/DCM), obtain 500mg, LCMS purity are 60%, and it is purified into (MP-A by preparation HPLC:10mM ammonium acetates (aqueous solution) MP-B：Acetonitrile Post：X bridge C18 (100x19) mm5 μ methods-T/%B:0/25/30,10/55, flow velocity：19ml/min eluant, eluents：THF+ ACN), (4S) -7- (2- picoline -4- bases)-N- (4- methylpyrimidine -2- bases) -3,4- dihydro-Isosorbide-5-Nitrae-endo-methylene group pyrroles are obtained Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (226mg, 0.582mmol, 36.7% yield), it is canescence Solid (TLC eluant, eluents：10%MeOH/EtOAc, R_f:0.4), LCMS (m/z):388.2[M+H]⁺。

¹H NMR(400MHz,CDCl3):δppm:13.74 (s, 1H), 8.63 (d, J=5.26Hz, 1H), 8.52 (d, J= 5.04Hz, 1H), 7.90-7.83 (m, 2H), 7.63 (m, J=8.11Hz, 1H), 7.47 (d, J=8.11Hz, 1H), 6.87 (d, J =5.04Hz, 1H), 5.78 (dd, J=5.92,3.29Hz, 1H), 3.32-3.10 (m, 3H), 3.10-2.88 (m, 1H), 2.68 (s,3H),2.54(s,3H),2.51-2.23(m,1H),2.23-1.99(m,1H)。

Embodiment 265

Synthesis (4S) -7- (1- methyl -2- oxo -1,2- dihydropyridine -4- bases)-N- (pyrazine -2- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature by potassium phosphate (939mg, 4.43mmol) and (1- methyl -2- oxo -1,2- dihydropyridine -4- bases) boric acid (462mg, 3.322mmol) adds to the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza- 5 (2H)-formamides (700mg, 2.215mmol) are in 1,4- dioxanes:Water (10mL, 8:2) mixture In solution in.By mixture purification for argon 30min.By PdOAc₂(49.62mg, 0.221mmol) and X-Phos (211.2mg, 0.443mmol) adds to reactant mixture, then stirs 16h at 100 DEG C.Reactant mixture is cooled to room temperature, It is concentrated in vacuo, by residue distribution between water (40mL) and EtOAc (80mL).Separate organic layer and through anhydrous Na₂SO₄Dry, Filter and evaporate filtrate, obtain crude compound.Crude compound is purified through column chromatography, using silica gel (100-200 mesh), 3% ethanol/methylene obtains (4S) -7- (1- methyl -2- oxo -1,2- dihydropyridine -4- bases)-N- (pyrroles as eluant, eluent Piperazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (110mg, 0.270mmol, 28.5% yield), it is faint yellow solid.(TLC eluant, eluents：10%MeOH/DCM R_f:0.4；UV is activated ), LCMS (m/z):390.30[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.53 (s, 1H), 9.53 (s, 1H), 8.34-8.28 (m, 2H), 7.63 (d, J=8.11Hz, 1H), 7.43 (t, J=7.78Hz, 2H), 7.20 (d, J=1.75Hz, 1H), 7.04 (dd, J=7.23, 1.97Hz, 1H), 5.70 (dd, J=5.92,3.07Hz, 1H), 3.62 (s, 3H), 3.33-3.14 (m, 3H), 3.03 (dd, J= 12.06,3.29Hz, 1H), 2.40-2.30 (m, 1H), 2.09 (dt, J=13.92,6.85Hz, 1H)

Embodiment 266

Synthesize (4S) -7- (5- cyano group -6- ethoxy pyridine -3- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines with argon-degassed 20min And [2,3-b] [1,4] diaza- 5 (2H)-formamides (400mg, 1.263mmol), 2- ethyoxyls -5- (4,4,5,5- tetramethyls Base -1,3,2- dioxaborolan -2- bases) pyridine -3- formonitrile HCNs (519mg, 1.894mmol) and K₃PO₄(536mg, X-phos (60.2mg, 0.126mmol), three 2.53mmol) are added in the solution in 1,4- dioxanes (15mL), water (3mL) (dibenzalacetone) two palladium (0) (57.8mg, 0.063mmol), uses argon-degassed 5min again.By reactant mixture 100 DEG C stirring 16 hours, be subsequently cooled to room temperature.Reactant mixture is filtered through diatomite, then filtrate water, which dilutes, is used in combination EtOAc extracts (3x20mL).Organic layer water, the aqueous salt solu-tion of merging, it is dried over sodium sulfate, and evaporated, obtain roughization Compound (TLC eluant, eluents：5%MeOH/DCM:R_f-0.3；UV activation).Crude compound is purified through column chromatography, uses neutral oxygen Change aluminium and use 30-40%EtOAc/ Hex, obtain pure (4S) -7- (5- cyano group -6- ethoxy pyridine -3- bases)-N- (pyrroles Piperazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (173mg, 0.398mmol, 31.5% yield), it is pale solid, LCMS (m/z):429.2[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 13.83 (s, 1H), 9.51 (d, J=1.5Hz, 1H), 9.16 (d, J=2.4Hz, 1H), 8.82 (d, J=2.6Hz, 1H), 8.55 (dd, J=2.6,1.5Hz, 1H), 8.31 (d, J=2.4Hz, 1H), 7.62 (d, J =7.9Hz, 1H), 7.38 (d, J=7.9Hz, 1H), 5.67 (dd, J=6.0,3.2Hz, 1H), 4.57 (q, J=7.1Hz, 2H), 3.12-3.32 (m, 3H), 3.01 (dd, J=12.0,3.3Hz, 1H), 2.34 (m, 1H), 2.07 (m, 1H), 1.48 (t, J= 7.1Hz,3H)。

Embodiment 267

Synthesize (4S) -7- (6- picoline -3- bases)-N- (tetrahydrochysene -2H- pyrans -4- bases) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature by triphosgene (212mg, 0.714mmol) slowly add in batches (4S) -7- (6- picoline -3- bases) - 2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(300mg, 1.190mmol) is in tetrahydrochysene furan In the solution for the stirring muttered in (THF) (15mL) and stirring 30min.By DIPEA (0.622mL, 3.57mmol) and tetrahydrochysene -2H- Pyrans -4- amine (180mg, 1.785mmol) adds to above-mentioned reactant mixture, and 16h is stirred at 70 DEG C.Reactant mixture is cooled to Room temperature, is concentrated in vacuo and by residue distribution between water (40mL) and EtOAc (80mL).Separate organic layer and through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into crude compound.Crude compound is purified through column chromatography, uses silica gel (100- 200 mesh), 2% ethanol/methylene obtains (4S) -7- (6- picoline -3- bases)-N- (tetrahydrochysene -2H- pyrroles as eluant, eluent Mutter -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (110mg, 0.290mmol, 29.2% yield), it is faint yellow solid.(TLC eluant, eluents：10%MeOH/DCM R_f:0.4；UV is activated ), LCMS (m/z):380.36[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 10.52 (br d, J=7.23Hz, 1H), 8.88 (d, J=1.97Hz, 1H), 7.92 (dd, J=8.11,2.41Hz, 1H), 7.54 (d, J=7.89Hz, 1H), 7.28 (s, 1H), 7.22 (d, J= 7.89Hz, 1H), 5.62 (dd, J=5.92,3.29Hz, 1H), 4.04-3.94 (m, 3H), 3.51 (tt, J=11.59, 2.33Hz, 2H), 3.27-3.15 (m, 2H), 3.13-3.06 (m, 1H), 2.95 (dd, J=11.84,3.29Hz, 1H) 2.63 (s, 3H), 2.26 (dddd, J=14.00,9.95,5.97,4.06Hz, 1H), 2.08-1.98 (m, 3H), 1.62-1.55 (m, 2H).

Embodiment 268

Synthesize (4S)-N- (Cvclopropvlmethvl) -7- (6- picoline -3- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaTEA is added in the solution of (200mg, 0.793mmol) in tetrahydrofuran (THF) (10mL) (0.331mL, 2.378mmol) and triphosgene (141mg, 0.476mmol), then stirs to room temperature, is kept for 30 minutes.Then plus Enter cyclopropyl-methylamine (85mg, 1.189mmol).Reactant mixture is stirred into 16h at 60 DEG C.So that reactant mixture reaches RT, And evaporate solvent, then use CH₂Cl₂(25mL) dilutes.By solution water and salt water washing, through anhydrous Na₂SO₄Dry, filtering And concentrate.Crude product is through flash column chromatography (silica gel：100-200 mesh, uses 5%MeOH/CH₂Cl₂Elution), obtain (4S)-N- (Cvclopropvlmethvl) -7- (6- picoline -3- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (140mg, 0.381mmol, 48.0% yield), it is faint yellow solid (TLC:R_f=0.3, only EtOAc), LCMS (m/z):350.30[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 10.52 (br s, 1H), 8.96 (d, J=1.97Hz, 1H), 8.02 (dd, J=8.00,2.30Hz, 1H), 7.53 (d, J=7.89Hz, 1H), 7.26-7.20 (m, 2H), 5.64 (dd, J=5.81, 3.18Hz, 1H), 3.32-3.05 (m, 5H), 2.95 (dd, J=11.84,3.29Hz, 1H), 2.62 (s, 3H), 2.26 (dddd, J =14.00,9.95,5.97,4.06Hz, 1H), 2.10-1.98 (m, 1H), 1.08 (dddt, J=12.78,7.66,5.04, 2.38,2.38Hz, 1H), 0.65-0.52 (m, 2H), 0.26 (q, J=4.97Hz, 2H).

Embodiment 269

(4S)-N- (3H- [1,2,3] triazol [4,5-d] pyrimidin-7-yl) -7- (2- picoline -4- bases) -3 are synthesized, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

By (4S)-N- (3H- [1,2,3] triazol [4,5-d] pyrimidin-7-yl) the chloro- 3,4- dihydros -1,4- bridge methylenes of -7- Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (250mg, 0.699mmol) and (2- picoline -4- bases) Solution of the boric acid (124mg, 0.908mmol) in 1,4- dioxanes (9mL) and water (1mL) is in 25 DEG C of degassing 15min.It is anti-to this Answer and Cs is added in mixture₂CO₃(455mg, 1.398mmol), then deaerate 15min again.It is eventually adding PdCl₂(dppf) (51.1mg, 0.070mmol) and reactant mixture is stirred into 2h at 100 DEG C.Reactant mixture is diluted with water into (10ml) to be used in combination Ethyl acetate extracts (2X10ml).The organic layer of merging is washed with water (10mL), salt solution (10ml), through anhydrous sodium sulfate drying simultaneously It is concentrated under reduced pressure, obtains crude compound.Through flash column chromatography (secondary), ((100-200 silica gel, uses 3%CH to crude product₂Cl₂/ MeOH is eluted), obtain compound.By its-cross preparation HPLC repurity, with 3% ethanol/methylene elute, obtain pure (4S)-N- (3H- [1,2,3] triazol [4,5-d] pyrimidin-7-yl) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (15mg, 0.036mmol, 5.15% yield), it is Pale solid.(TLC systems:5% methanol/DCM.R_fValue：0.3), LCMS (m/z):414.9[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.23 (br s, 1H), 8.47 (br d, J=19.95Hz, 2H), 8.08(br s,1H),7.92-8.04(m,1H),7.67-7.81(m,2H),5.48-5.64(m,1H),3.20-3.26(m, 1H), 3.15 (br d, J=11.40Hz, 2H), 2.90-3.05 (m, 1H), 2.46 (br s, 3H), 2.20-2.38 (m, 1H), 1.92-2.09(m,1H)。

Embodiment 270

Synthesize (4S)-N- (3- Calmazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaTriethylamine is added in the solution of the stirring of (500mg, 1.982mmol) in THF (10mL, seal pipe) (1.657mL, 11.89mmol) and triphosgene (588mg, 1.982mmol) and stirring 30min, then add 3- Calmazine -2- amine (336mg, 2.97mmol), then heats 15h at 80 DEG C.Reactant mixture is cooled to room temperature, then distributed at water (25mL) Between EtOAc (40mL).Separate organic layer and through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain rough chemical combination Thing.Crude compound is through flash column chromatography (silica gel:100-200 mesh, eluant, eluent：2% methanol/DCM), obtain (4S)-N- (3- Calmazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine- 5 (2H)-formamides (27mg, 0.068mmol, 3.44% yield), it is pale solid (TLC eluant, eluents：5% first Alcohol/DCM, R_f:0.4), LCMS (m/z):392.3[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.43 (s, 1H), 8.59 (d, J=5.26Hz, 1H), 8.32 (d, J= 2.63Hz, 1H), 7.90 (d, J=2.41Hz, 1H), 7.72-7.55 (m, 3H), 7.37 (br d, J=4.38Hz, 1H), 5.73 (br dd, J=5.70,3.07Hz, 1H), 3.36-3.02 (m, 4H), 2.61 (m, 3H), 2.33 (br dd, J=9.87, 4.17Hz, 1H), 2.12 (br dd, J=9.72,3.86Hz, 1H).

Embodiment 271

Synthesize (4S)-N- (pyridine -2- bases) -7- (3- (trifluoromethyl) pyrrolidin-1-yl) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides)

At 0 DEG C to (4S) -7- (3- (trifluoromethyl) pyrrolidin-1-yl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza(peak 2, the SFC separation from intermediate) (350mg, 1.173mmol) is in tetrahydrofuran (THF) triphosgene (209mg, 0.704mmol) is added in the solution in (10mL).After 30min add DIPEA (1.025mL, 5.87mmol) with pyridine -2- amine (221mg, 2.347mmol), reactant mixture is then stirred into 16h at 70 DEG C.Reaction is mixed Compound is poured on cold water (50mL) and is extracted with ethyl acetate (2x50mL).Organic layer is concentrated under reduced pressure through anhydrous sodium sulfate drying, Obtain crude compound.Crude product is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：2%MeOH/DCM), obtain (4S)-N- (pyridine -2- bases) -7- (3- (trifluoromethyl) pyrrolidin-1-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza- 5 (2H)-formamides (50.8mg, 0.115mmol, 9.83% yield), it is faint yellow solid. (TLC systems:R_f:02,:EtOAc), LCMS (m/z):419.2[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δppm 13.34(s,1H),8.25-8.11(m,2H),7.69-7.61(m,1H), 7.34 (d, J=8.33Hz, 1H), 6.98-6.90 (m, 1H), 5.98 (d, J=8.55Hz, 1H), 5.61 (dd, J=6.14, 3.29Hz, 1H), 4.02 (dd, J=11.18,8.55Hz, 1H), 3.82 (dd, J=11.29,7.34Hz, 1H), 3.70 (td, J =8.93,4.28Hz, 1H), 3.63-3.51 (m, 1H), 3.29-3.06 (m, 4H), 2.90 (dd, J=11.84,3.29Hz, 1H),2.39-2.19(m,3H),2.05-1.91(m,1H)。

Embodiment 272

Synthesize (4S) -7- (2- picoline -4- bases)-N- (9H- purine -6- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diazaIn the solution of (1g, 3.96mmol) in tetrahydrofuran (THF) (20mL) add triphosgene (0.588g, 1.982mmol) and stirring 1h.Add triethylamine (2.76mL, 19.82mmol), be subsequently added into 9H- purine -6- amine (0.696g, 5.15mmol).Reactant mixture is stirred into 16h at 70 DEG C.Reactant mixture is diluted with water and uses 2x25ml ethyl acetate to extract Take.The organic layer aqueous salt solu-tion of merging, through anhydrous sodium sulfate drying and is concentrated under reduced pressure, and obtains crude compound.Thick production Through flash column chromatography, (100-200 silica gel, uses 3%CH to thing₂Cl₂/ MeOH is eluted), desired compound is obtained, is passed through Preparation HPLC is purified, and obtains (4S) -7- (2- picoline -4- bases)-N- (9H- purine -6- bases) -3,4- dihydros-Isosorbide-5-Nitrae-bridge Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (25mg, 0.057mmol, 1.449% yield), its For white solid.(TLC systems:5% methanol/DCM.R_fValue：0.3), LCMS (m/z):411.9[M+H]⁺。

¹H NMR(400MHz,DMSO-d₆):δppm 14.08(br s,1H),12.32(br s,1H),8.72(s,1H), 8.63 (d, J=5.04Hz, 1H), 8.45 (s, 1H), 8.20 (br s, 1H), 7.99 (dd, J=5.48,1.53Hz, 1H), 7.90-7.81 (m, 1H), 7.81-7.74 (m, 1H), 5.57 (dd, J=5.70,3.07Hz, 1H), 3.27-3.07 (m, 3H), 3.02 (br dd, J=11.95,3.18Hz, 1H), 2.65 (s, 3H), 2.37-2.18 (m, 1H), 2.08 (br d, J= 6.58Hz,1H)。

Embodiment 273

Synthesize (4S) -7- (6- picoline -3- bases)-N- (2,2,2- trifluoroethyls) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Triphosgene (212mg, 0.74mmol) is slowly added into (4S) -7- (6- picoline -3- bases) -2 in batches in room temperature, 3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(300mg, 1.190mmol) is in tetrahydrofuran (THF) in the solution of the stirring in (15mL) and stirring 30min.By DIPEA (0.622mL, 3.571mmol) and 2,2,2- trifluoros Ethylamine hydrochloride (242mg, 1.785mmol) adds to above-mentioned reactant mixture, and 16h is stirred at 70 DEG C.Reactant mixture is cooled down To room temperature, it is concentrated in vacuo, by residue distribution between water (40mL) and EtOAc (80mL).Separate organic layer and through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into crude compound.Crude compound is purified through column chromatography, uses silica gel (100- 200 mesh), 2% ethanol/methylene obtains (4S) -7- (6- picoline -3- bases)-N- (2,2,2- trifluoros as eluant, eluent Ethyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (235mg, 0.615mmol, 103% yield), it is faint yellow solid.(TLC eluant, eluents：10%MeOH/DCM R_f:0.5；UV activation), LCMS(m/z)378.31[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 11.12 (br t, J=6.03Hz, 1H), 8.88 (d, J=1.97Hz, 1H), 7.94 (dd, J=8.11,2.41Hz, 1H), 7.57 (d, J=7.89Hz, 1H), 7.31-7.22 (m, 2H), 5.62 (dd, J =6.03,3.18Hz, 1H), 4.16-4.02 (m, 2H), 3.32-3.08 (m, 3H), 2.96 (dd, J=12.06,3.29Hz, 1H), 2.62 (s, 3H), 2.28 (dddd, J=14.06,10.00,6.08,4.06Hz, 1H), 2.09-1.97 (m, 1H)

Embodiment 274

Synthesize (4S) -7- (6- picoline -3- bases)-N- (tetrahydrochysene -2H- pyrans -3- bases) -3,4- dihydro 1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaTriphosgene is added in the solution of (300mg, 1.189mmol) in tetrahydrofuran (THF) (10mL) (176mg,0.594mmol).Addition DIPEA (1.038mL, 5.94mmol) after 30min, tetrahydrochysene -2H- pyrans -3- amine (180mg, 1.783mmol) and by reactant mixture at 70 DEG C stir 16h.Reactant mixture is poured on cold water (50mL) and acetic acid second is used Ester extracts (150mL).Organic layer is concentrated under reduced pressure through anhydrous sodium sulfate drying, obtains crude compound.Crude product is through quick post color Spectrum purifying (silica gel：100-200 mesh, eluant, eluent：3%MeOH/DCM), (4S) -7- (6- picoline -3- bases)-N- (four is obtained Hydrogen -2H- pyrans -3- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (114.5mg, 0.299mmol, 25.1% yield), it is pale solid.(TLC systems:R_f:0.2,5%MeOH-DCM), LCMS(m/z):380.0[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 10.59 (t, J=7.78Hz, 1H), 8.92 (s, 1H), 8.07 (dt, J= 8.11,2.96Hz, 1H), 7.52 (d, J=7.89Hz, 1H), 7.27-7.17 (m, 2H), 5.66-5.59 (m, 1H), 4.10- 4.00 (m, 1H), 3.86 (dt, J=11.24,3.70Hz, 1H), 3.71-3.62 (m, 2H), 3.53 (dd, J=11.18, 5.70Hz, 1H), 3.28-3.06 (m, 3H), 2.97-2.89 (m, 1H), 2.60 (s, 3H), 2.24 (dddd, J=16.66, 9.98,6.25,3.51Hz,1H),2.07-1.91(m,2H),1.74-1.66(m,2H),1.56-1.50(m,1H)。

Embodiment 275

Synthesize (4S) -7- ((R) -2- ethyl morpholine generations)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

Diisopropylethylamine (2.81mL, 16.12mmol) is added into pyrazine -2- formic acid at 30 DEG C under argon gas stirring In the solution of the stirring of (400mg, 3.22mmol) in tetrahydrofuran (15mL).By diphenyl phosphate azide (887mg, Reactant mixture 3.22mmol) is added to, then 2h is stirred at 30 DEG C.Then by (2R) -2- ethyls -4- ((4S) -2,3,4,5- four Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 7- bases) morpholine (619mg, 2.256mmol) adds to reaction Mixture and 65 DEG C stir 16h.By reactant mixture be cooled to room temperature and distribute water (20mL) and ethyl acetate (2 × Between 30mL).Organic layer is separated, then through anhydrous sodium sulfate drying, filters and evaporates filtrate, obtain rough thing.(TLC is washed De- agent：90% ethyl acetate/hexane, R_f=0.3；UV activation).Thick material is purified through column chromatography, using neutral alumina, will Product is eluted with 10-15% ethyl acetate/hexanes, obtains pure (4S) -7- ((R) -2- ethyl morpholine generations)-N- (pyrazine -2- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (35mg, 0.088mmol, 2.75% yield), it is pale solid, LCMS (m/z):396.31[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 13.37 (s, 1H), 9.55 (d, J=1.53Hz, 1H), 8.25 (d, J= 2.63Hz, 1H), 8.09-8.18 (m, 1H), 7.38 (d, J=8.55Hz, 1H), 6.30 (d, J=8.55Hz, 1H), 5.63 (dd, J=6.03,3.18Hz, 1H), 4.00-4.13 (m, 2H), 3.86 (br d, J=12.93Hz, 1H), 3.75 (td, J=11.62, 2.85Hz, 1H), 3.45-3.56 (m, 1H), 3.13-3.30 (m, 1H), 3.00-3.17 (m, 3H), 2.92 (dd, J=11.84, 3.29Hz, 1H), 2.69-2.81 (m, 1H), 2.27 (dddd, J=13.89,9.95,6.08,3.62Hz, 1H), 2.00 (dt, J =14.09,6.88Hz, 1H), 1.59-1.78 (m, 2H), 1.05 (t, J=7.45Hz, 3H).

Embodiment 276

Synthesize (4S) -7- (3- methyl isophthalic acid H- pyrazoles -5- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Tripotassium phosphate (469mg, 2.210mmol) is added into the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- two at 28 DEG C Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (350.0mg, 1.105mmol), 3- Methyl -5- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) -1H- pyrazoles (276mg, 1.326mmol) exists In 1,4- dioxanes (10.0ml) and the solution of the stirring in water (2.0ml).Reactant mixture is deaerated 15min, thereto plus Enter X-phos (52.7mg, 0.110mmol) and Pd₂(dba)₃(50.6mg,0.055mmol).Reactant mixture is deaerated again 15min is simultaneously stirred 16 hours at 100 DEG C.Then mixture is cooled to 28 DEG C, is then filtered by Celite pad and use acetic acid Ethyl ester washs (40mL).Filtrate water washs (10mL).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and by filtrate Evaporation, obtains thick material, and it is brown solid (TLC eluant, eluents：10%MeOH/DCM；R_f-0.3；UV activation).By the thick thing Matter purifies through GRACE posts and uses 2-5%MeOH/ dichloromethane eluents, obtains pure (4S) -7- (3- methyl isophthalic acid H- pyrazoles -5- Base)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine (181.5mg, 0.492mmol, 44.5% yield), it is white solid, LCMS (m/z):363.27[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 13.80 (s, 1H), 9.55 (d, J=1.10Hz, 1H), 8.52 (s, 1H), 8.20-8.35 (m, 2H), 7.55 (d, J=8.11Hz, 1H), 7.12 (d, J=7.89Hz, 1H), 5.68 (dd, J=5.92, 3.29Hz, 1H), 3.11-3.34 (m, 3H), 2.89-3.11 (m, 1H), 2.60 (s, 3H), 2.33 (dddd, J=14.06, 9.95,5.92,4.06Hz,1H),1.97-2.17(m,1H)

Embodiment 277

Synthesis (4S)-N- isopropyls -7- (6- picoline -3- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza- 5 (2H)-formamides

0 DEG C of priority by DIPEA (1.038mL, 5.94mmol) and triphosgene (353mg, 1.189mmol) add to (4S)- 7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza In the solution of the stirring of (500mg, 1.982mmol) in THF (15mL, in seal pipe), 30min is then stirred at room temperature. Then propyl- 2- amine (176mg, 2.97mmol) is added, 75 DEG C are then heated to, 16h is kept.Reactant mixture is cooled to room temperature And distribute between water (25mL) and EtOAc (60mL).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and by filtrate Evaporation, obtains crude compound.Crude mixture is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：2%MeOH/ DCM), (4S)-N- isopropyls -7- (6- picoline -3- bases) -3,4- dihydro-Isosorbide-5-Nitrae-endo-methylene group pyridos [2,3-b] are obtained [1,4] diaza- 5 (2H)-formamides (130mg, 0.384mmol, 19.39% yield), it is that (TLC is washed pale solid De- agent：5%MeOH/DCM, R_f:0.3), LCMS (m/z):338.32[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 10.38 (d, J=6.4Hz, 1H), 8.91 (d, J=2Hz, 1H), 7.94 (dd, J=2.4,8Hz, 1H), 7.52 (d, J=5.6Hz, 1H), 7.25-7.20 (m, 2H), 5.66-5.62 (m, 1H), 4.14- 4.06(m,1H),3.26-3.06(m,3H),2.97-2.92(m,1H),2.62(s,3H),2.30-2.21(m,1H),2.26- 1.98 (m, 1H), 1.27 (d, J=6.8Hz, 6H)

Embodiment 278

Synthesize (4S)-N- (1- methyl piperidine -4- bases) -7- (6- picoline -3- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Triphosgene (423.8mg, 1.428mmol) is slowly added into (4S) -7- (6- picolines -3- in batches in room temperature Base) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(600mg, 2.380mmol) is four In the solution of stirring in hydrogen furans (THF) (25mL) and stirring 30min.By DIPEA (1.24mL, 7.14mmol) and 1- methyl Piperidines -4- amine (408mg, 3.571mmol) adds to reactant mixture, and 16h is stirred at 70 DEG C.Reactant mixture is cooled to room Temperature, is concentrated in vacuo, by residue distribution between water (40mL) and EtOAc (80mL).Separate organic layer and through anhydrous Na₂SO₄It is dry It is dry, filter and filtrate evaporation is obtained into crude compound.Crude compound is purified through column chromatography, using silica gel (100-200 mesh), 2% ethanol/methylene as eluant, eluent, obtain (4S)-N- (1- methyl piperidine -4- bases) -7- (6- picoline -3- bases) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (100mg, 0.252mmol, 21.21% yield), it is pale solid, and it is faint yellow solid.(TLC eluant, eluents：10%MeOH/DCM R_f:0.4；UV Activation), LCMS (m/z):393.33[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ ppm 10.46 (br d, J=7.45Hz, 1H), 8.88 (d, J=2.19Hz, 1H), 7.92 (dd, J=8.11,2.41Hz, 1H), 7.53 (d, J=7.89Hz, 1H), 7.29-7.24 (m, 1H), 7.20 (d, J =7.89Hz, 1H), 5.62 (dd, J=5.92,3.29Hz, 1H), 3.82-3.73 (m, 1H), 3.12-3.26 (m, 2H), 3.12- 3.06 (m, 1H), 2.94 (dd, J=11.84,3.29Hz, 1H), 2.85-2.78 (m, 1H), 2.86-2.77 (m, 1H), 2.62 (s,3H),2.30-2.22(m,4H),2.15-1.98(m,5H),1.65-1.53(m,2H)。

Embodiment 279

Synthesize (4S) -7- ((2R, 6R) -2,6- thebaine generations)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Diphenyl phosphate azide (0.432g, 1.571mmol) is added into pyrazine -2- formic acid in room temperature in argon gas atmosphere In the solution of the stirring of (0.195g, 1.571mmol) and DIPEA (1.372mL, 7.86mmol) in tetrahydrofuran (30mL). 2h is stirred at room temperature in reactant mixture, be subsequently added (2R, 6R) -2,6- dimethyl -4- ((4S) -2,3,4,5- tetrahydrochysenes-Isosorbide-5-Nitrae - Endo-methylene group pyrido [2,3-b] [1,4] diaza- 7- bases) morpholine (0.302g, 1.100mmol) and by reactant mixture 16h is stirred at 65 DEG C.Reactant mixture is cooled to room temperature and evaporating completely solvent is depressurized, then distribute in water (20mL) and Between EtOAc (60mL).Organic layer is separated, through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain thick material, it is palm fibre Color solid (TLC eluant, eluents：100%EtOAc:R_f-0.3；UV activation).Thick material is purified through column chromatography, uses neutral alumina Aluminium, and 20%EtOAc/ Hex is used, obtain pure (4S) -7- ((2R, 6R) -2,6- thebaine generations)-N- (pyrroles Piperazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (0.081g, 0.203mmol, 12.93% yield), it is white solid, LCMS (m/z):396.38[M+H]⁺。

¹H NMR (400MHz, cdcl3-d) δ ppm 13.42 (s, 1H), 9.56 (d, J=1.32Hz, 1H), 8.25 (d, J =2.63Hz, 1H), 8.09-8.19 (m, 1H), 7.37 (d, J=8.55Hz, 1H), 6.24 (d, J=8.55Hz, 1H), 5.63 (dd, J=6.03,3.18Hz, 1H), 4.28-4.15 (m, 2H), 3.67 (dd, J=12.39,3.40Hz, 2H), 3.32-3.19 (m, 3H), 3.17-3.06 (m, 2H), 2.92 (dd, J=11.84,3.29Hz, 1H), 2.27 (dddd, J=13.95,10.00, 6.19,3.84Hz, 1H), 2.00 (dt, J=14.09,7.10Hz, 1H), 1.34 (d, J=6.36Hz, 6H).

Embodiment 280

Synthesize (4S) -7- (1- (difluoromethyl) -1H- pyrazoles -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Tripotassium phosphate (469mg, 2.210mmol) is added into the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (350mg, 1.105mmol), 1- (difluoro first Base) -4- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) -1H- pyrazoles (324mg, 1.326mmol) 1, In 4- dioxanes (10.0ml) and the solution of the stirring in water (2.0ml).Reactant mixture is deaerated 10min.By Pd₂(dba)₃ (50.6mg, 0.055mmol) and X-phos (52.7mg, 0.110mmol) add to reactant mixture, and then deaerate 15min again.Will Reactant mixture is stirred 16 hours at 100 DEG C.Reactant mixture is cooled to 28 DEG C, is then filtered by Celite pad and uses second Acetoacetic ester washs (40mL).Filtrate water washs (10ml).Organic layer is separated, then through anhydrous Na₂SO₄Dry, filter and will filter Liquid evaporates, and obtains thick material, and it is brown solid (TLC eluant, eluents：10%MeOH/DCM:R_f-0.4；UV activation).By thick thing Matter is purified through GRACE posts, and uses 1-2%MeOH/ dichloromethane eluents, obtains pure (4S) -7- (1- (difluoromethyl) -1H- Pyrazoles -4- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (280.0mg, 0.702mmol, 63.6% yield), is white solid, LCMS (m/z):399.30[M+H]⁺。

¹H NMR(400MHz,CDCl₃):δ 14.18 (s, 1H), 9.55 (d, J=1.3Hz, 1H), 9.17 (s, 1H), 8.36- 8.27 (m, 2H), 8.22 (s, 1H), 7.56 (d, J=7.9Hz, 1H), 7.26-7.06 (m, 2H), 5.66 (dd, J=5.9, 3.3Hz, 1H), 3.35-3.11 (m, 3H), 3.01 (dd, J=12.3,3.3Hz, 1H), 2.34 (dddd, J=14.1,9.8, 6.0,4.06Hz,1H),1.93-2.17(m,1H)。

Embodiment 281

(4S) -7- (6- (dimethylamino) -4- picoline -3- bases)-N- (pyridine -2- bases) -3,4- dihydro -1 is synthesized, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature successively by N, N, 4- trimethyls -5- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) Pyridine -2- amine (274mg, 1.045mmol) and K₃PO₄(605mg, 2.85mmol) add to the chloro- N- of (4S) -7- (pyridine -2- bases) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (300mg, 0.950mmol) In the solution of stirring in n-butyl alcohol (10mL) and the 20min that deaerates.Then by Pd₂(dba)₃(43.5mg, 0.048mmol) and X-phos (45.3mg, 0.095mmol) adds to reactant mixture and reactant mixture is stirred into 16h at 80 DEG C.By reaction mixing Thing is cooled to room temperature, then distributes between water (25mL) and EtOAc (40mL).Separate organic layer and through anhydrous Na₂SO₄Dry, Filter and filtrate evaporation is obtained into crude compound.Crude compound purifies (silica gel through column chromatography:100-200 mesh), by chemical combination Thing is collected in 70%EtOAc/ petroleum ethers, obtains (4S) -7- (6- (dimethylamino) -4- picoline -3- bases)-N- (pyrroles Pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (100mg, 0.236mmol, 24.80% yield), it is pale solid.(TLC eluant, eluents：100%EtOAc/ petroleum ethers R_f:0.4, UV lives Change), LCMS (m/z):416.3[M+H]⁺。

¹H NMR(400MHz,CDCl₃)δppm 13.31(s,1H),8.36-8.17(m,3H),8.17-7.96(m,1H), 7.86-7.57 (m, 1H), 6.99 (d, J=7.89Hz, 1H), 6.95-6.68 (m, 1H), 6.43 (s, 1H), 5.69 (brdd, J= 5.59,3.18Hz, 1H), 3.31 (br d, J=8.77Hz, 1H), 3.24-3.10 (m, 8H), 3.10-2.87 (m, 1H), 2.47 (s,3H),2.40-2.20(m,1H),2.19-1.92(m,1H)

Embodiment 282

Synthesize (4S) -7- (6- (piperidin-1-yl) pyridin-3-yl)-N- (pyridine -2- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature by (6- (piperidin-1-yl) pyridin-3-yl) boric acid (215mg, 1.045mmol) successively will and K₃PO₄ (605mg, 2.85mmol) adds to the chloro- N- of (4S) -7- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaIn the solution of stirring of -5 (the 2H)-formamides (300mg, 0.950mmol) in n-butyl alcohol (10mL) simultaneously Deaerate 20min.Then by Pd₂(dba)₃(43.5mg, 0.048mmol) and X-phos (45.3mg, 0.095mmol) add to reaction Reactant mixture is simultaneously stirred 16h by mixture at 80 DEG C.Reactant mixture is cooled to room temperature, then distribute in water (25mL) and Between EtOAc (40mL).Separate organic layer and through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into crude compound.Slightly Produced compounds purify (silica gel through column chromatography:100-200 mesh), compound is collected in 70%EtOAc/ petroleum ethers, obtained (4S) -7- (6- (piperidin-1-yl) pyridin-3-yl)-N- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza- 5 (2H)-formamides (150mg, 0.336mmol, 35.3% yield), it is faint yellow solid. (TLC eluant, eluents：100%EtOAc/ petroleum ethers, R_f:0.4, UV activation), LCMS (m/z):442.3[M+H]⁺。

¹H NMR (400MHz, CDCl3) δ ppm 13.71 (s, 1H), 8.80 (d, J=2.63Hz, 1H), 8.54 (dd, J= 8.99,2.63Hz, 1H), 8.37 (dt, J=4.93,0.82Hz, 1H), 8.18 (d, J=8.33Hz, 1H), 7.78-7.59 (m, 1H), 7.51 (d, J=8.11Hz, 1H), 7.30-7.25 (m, 1H), 6.97 (ddd, J=7.23,4.82,0.88Hz, 1H), 6.79 (d, J=8.99Hz, 1H), 5.68 (dd, J=5.92,3.29Hz, 1H), 3.70-3.59 (m, 4H), 3.09-3.32 (m, 3H), 3.09-2.84 (m, 1H), 2.31 (d, J=1.97Hz, 1H), 2.17-1.95 (m, 1H), 1.68 (br s, 6H).

Embodiment 283

Synthesize (4S)-N- (tert-butyl group) -7- (6- picoline -3- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature successively by DIPEA (1.038mL, 5.94mmol) and triphosgene (353mg, 1.189mmol) add to (4S)- 7- (6- picoline -3- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza In the solution of (500mg, 1.982mmol) in tetrahydrofuran (15mL), 30min is then stirred for.Then 2- methyl is added Propyl- 2- amine (217mg, 2.97mmol), is then heated to 75 DEG C, keeps 16h.Reactant mixture is cooled to 28 DEG C, Ran Houfen Fit between water (25mL) and EtOAc (60mL).The organic layer of separation is through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain Obtain crude compound.Crude mixture is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：2%MeOH/DCM), obtain To (4S)-N- (tert-butyl group) -7- (6- picoline -3- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] Diaza- 5 (2H)-formamides (185mg, 0.525mmol, 26.5% yield), it is pale solid (TLC eluant, eluents： 5%MeOH/DCM, R_f:0.3；UV activation), LCMS (m/z):352.34[M+H]⁺。

¹H NMR(400MHz,CDCl₃-d):δ ppm 10.4 (s, 1H), 8.90 (d, J=2Hz, 1H), 7.95 (dd, J= 2.4Hz, 8Hz, 1H), 7.51 (d, J=7.6Hz, 1H), 7.22 (d, J=8Hz, 1H), 7.17 (d, J=8Hz, 1H), 5.64- 5.61(m,1H),3.28-3.12(m,2H),3.10-3.05(m,1H),2.96-2.91(m,1H),2.61(s,3H),2.30- 2.21 (m, 1H), 2.06-1.97 (m, 1H), 1.43 (s, 9H).

Embodiment 284

Synthesize (4S)-N- (pyridine -2- bases) -7- (2- (pyrrolidin-1-yl) pyrimidine -5- bases) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature successively by 2- (pyrrolidin-1-yl) -5- (4,4,5,5- tetramethyl -1,3,2- dioxaborolans -2- Base) pyrimidine (288mg, 1.045mmol) and K₃PO₄(605mg, 2.85mmol) adds to the chloro- N- of (4S) -7- (pyridine -2- bases) -3, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (300mg, 0.950mmol) exist In the solution of stirring in n-butyl alcohol (10mL) and the 20min that deaerates.Then by Pd₂(dba)₃(43.5mg, 0.048mmol) and X- Phos (45.3mg, 0.095mmol) adds to reactant mixture and reactant mixture is stirred into 16h at 80 DEG C.By reactant mixture Room temperature is cooled to, is then distributed between water (25mL) and EtOAc (40mL).Separate organic layer and through anhydrous Na₂SO₄Dry, mistake Filter and filtrate evaporation is obtained into crude compound.Crude compound purifies (silica gel through column chromatography:100-200 mesh), by compound Collected in 70%EtOAc/ petroleum ethers, acquisition (4S)-N- (pyridine -2- bases) -7- (2- (pyrrolidin-1-yl) pyrimidine -5- bases) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (170mg, 0.377mmol, 39.7% yield), it is yellow solid.(TLC eluant, eluents：100%EtOAc/ petroleum ethers, R_f:0.4, UV activation), LCMS (m/z):429.6[M+H]⁺。

¹H NMR(400MHz,CDCl₃)δppm 13.57(s,1H),9.09(s,2H),8.51-8.26(m,1H),8.14 (d, J=8.33Hz, 1H), 7.67 (td, J=7.84,1.86Hz, 1H), 7.54 (m, J=8.11Hz, 1H), 7.26 (s, 1H), 6.97 (ddd, J=7.29,4.88,0.99Hz, 1H), 5.68 (dd, J=5.81,3.18Hz, 1H), 3.75-3.60 (m, 4H), 3.35-3.09 (m, 3H), 2.99 (dd, J=12.06,3.29Hz, 1H), 2.32 (m, 1H), 1.98-2.13 (m, 5H)

Embodiment 285

Synthesize (4S) -7- (6- (diethylamino) pyridin-3-yl)-N- (pyridine -2- bases) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature successively by N, N- diethyl -5- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) pyrrole Pyridine -2- amine (289mg, 1.045mmol) and K₃PO₄(605mg, 2.85mmol) adds to the chloro- N- of (4S) -7- (pyridine -2- bases) -3, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (300mg, 0.950mmol) exist In the solution of stirring in n-butyl alcohol (10mL) and the 20min that deaerates.Then by Pd₂(dba)₃(43.5mg, 0.048mmol) and X- Phos (45.3mg, 0.095mmol) adds to reactant mixture and reactant mixture is stirred into 16h at 80 DEG C.By reactant mixture Room temperature is cooled to, is then distributed between water (25mL) and EtOAc (40mL).Separate organic layer and through anhydrous Na₂SO₄Dry, mistake Filter and filtrate evaporation is obtained into crude compound.Crude compound purifies (silica gel through column chromatography:100-200 mesh), by compound Collected in 70%EtOAc/ petroleum ethers, acquisition (4S) -7- (6- (diethylamino) pyridin-3-yl)-N- (pyridine -2- bases) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (120mg, 0.271mmol, 28.5% yield), it is pale solid.(TLC eluant, eluents：100%EtOAc/ petroleum ethers, R_f:0.4, UV activation), LCMS (m/z):430.3[M+H]⁺。

¹H NMR(400MHz,CDCl₃) δ ppm 13.77 (s, 1H), 8.78 (d, J=2.41Hz, 1H), 8.64-8.43 (m, 1H), 8.43-8.28 (m, 1H), 8.18 (d, J=8.55Hz, 1H), 7.75-7.59 (m, 1H), 7.51 (d, J=8.11Hz, 1H), 7.39-7.15 (m, 1H), 6.97 (ddd, J=7.34,4.93,0.88Hz, 1H), 6.62 (d, J=8.99Hz, 1H), 5.68 (dd, J=5.92,3.07Hz, 1H), 3.69-3.50 (m, 4H), 3.34-3.08 (m, 3H), 3.08-2.89 (m, 1H), 2.39-2.19 (m, 1H), 2.08 (dt, J=13.87,6.77Hz, 1H), 1.24 (t, J=7.02Hz, 6H)

Embodiment 286

Synthesize (4S) -7- (6- morpholinoes pyridin-3-yl)-N- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature successively by (6- morpholinoes pyridin-3-yl) boric acid (296mg, 1.425mmol) and K₃PO₄(605mg, 2.85mmol) add to the chloro- N- of (4S) -7- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] DiazaIn the solution of stirring of -5 (the 2H)-formamides (300mg, 0.950mmol) in n-butyl alcohol (10mL) and deaerate 20min.Then by Pd₂(dba)₃(43.5mg, 0.048mmol) and X-phos (45.3mg, 0.095mmol) add to reaction mixing Reactant mixture is simultaneously stirred 16h by thing at 80 DEG C.Reactant mixture is cooled to room temperature, then distribute in water (25mL) and Between EtOAc (40mL).Separate organic layer and through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into crude compound.Slightly Produced compounds purify (silica gel through column chromatography:100-200 mesh), compound is collected in 70%EtOAc/ petroleum ethers, obtained (4S) -7- (6- morpholinoes pyridin-3-yl)-N- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (110mg, 0.245mmol, 25.8% yield), it is pale solid.(TLC is washed De- agent：100%EtOAc/ petroleum ethers, R_f:0.4, UV activation), LCMS (m/z):444.3[M+H]⁺。

¹H NMR(400MHz,CDCl₃-d)δppm 13.68(s,1H),8.85(s,1H),8.56-8.64(m,1H),8.36 (d, J=3.73Hz, 1H), 8.19 (d, J=8.33Hz, 1H), 7.68 (t, J=7.13Hz, 1H), 7.54 (d, J=8.11Hz, 1H), 7.32 (s, 1H), 7.02-6.95 (m, 1H), 6.78 (d, J=8.99Hz, 1H), 5.68 (dd, J=5.70,3.07Hz, 1H), 3.91-3.83 (m, 4H), 3.70-3.60 (m, 4H), 3.34-3.14 (m, 3H), 2.99 (dd, J=11.95,3.18Hz, 1H), 2.42-2.26 (m, 1H), 2.08 (dt, J=13.87,7.21Hz, 1H).

Embodiment 287

Synthesize (4S)-N- (pyrazine -2- bases) -7- (2H- tetrazolium -5- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides

Azido tributylstannyl (1.297g, 3.90mmol) is added into (4S) -7- cyano group-N- (pyrazine -2- bases) -3, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (0.4g, 1.302mmol) exist In the solution of stirring in tetrahydrofuran (25mL), 80-90 DEG C is then heated in seal pipe, 48h is kept.By reaction mixing Thing is cooled to room temperature.Tetrahydrofuran is concentrated under reduced pressure, residue is obtained.By residue with dchloromethane (100mL).To have Machine layer 50mL 0.5M HCl, water (50mL) and salt solution (50mL) washing.Organic layer is filtered and steamed through anhydrous sodium sulfate drying Hair, obtains thick material (TLC eluant, eluents：10%MeOH/DCM；UV is activated；R_f~0.3).Crude compound triturated under ether and mistake Filter, obtains (4S)-N- (pyrazine -2- bases) -7- (2H- tetrazolium -5- bases) -3,4- dihydro-Isosorbide-5-Nitrae-endo-methylene group pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (0.13g, 0.369mmol, 28.4% yield), it is white solid, LCMS (m/ z):351.24[M+H]⁺。

¹H NMR(400MHz,CDCl₃- d) δ ppm 14.75 (br s, 1H), 9.47 (d, J=1.32Hz, 1H), 8.48- 8.40 (m, 2H), 8.02 (d, J=7.89Hz, 1H), 7.77 (d, J=7.89Hz, 1H), 5.57 (dd, J=5.92,3.07Hz, 1H), 3.37-3.27 (m, 2H), 3.22-3.17 (m, 1H), 3.24-3.04 (m, 1H), 2.39 (ddt, J=14.36,8.88, 5.48,5.48Hz,1H),2.18-2.05(m,1H)

Embodiment 288

Synthesize (4S)-N- (1H- benzos [d] imidazoles -5- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diazaEt is added in the solution of (300mg, 0.983mmol) in THF (20mL)₃N(0.822mL, 5.90mmol), triphosgene (292mg, 0.983mmol) and stirring 1h.Then add 1H- benzos [d] imidazoles -5- amine (393mg, 2.95mmol) and by reactant mixture at 65 DEG C heat 15h.(TLC eluant, eluents：100%EtOAc:R_f-0.2；UV activation). Reactant mixture is cooled to room temperature, is concentrated in vacuo and by residue distribution between water (30mL) and DCM (50mL).Separation has Machine layer simultaneously obtains crude compound through anhydrous sodium sulfate drying, filtering and by filtrate evaporation.Crude product is through flash column chromatography (neutral alumina, eluant, eluent：70% ethyl acetate/hexane), obtain desired product (4S)-N- (1H- benzos [d] imidazoles -5- Base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (215mg, 0.461mmol, 47.0% yield), it is pale solid.LCMS(m/z):465.09[M+H ]⁺,R_t=1.77min.

¹H NMR(400MHz,DMSO-d₆):δppm 13.02-12.63(m,1H),12.34(br s,1H),8.30-8.20 (m,2H),8.16-8.10(m,1H),8.06(s,1H),7.92-7.77(m,2H),7.73-7.67(m,1H),7.67-7.60 (m, 1H), 7.52-7.37 (m, 1H), 7.18-6.91 (m, 1H), 5.53 (dd, J=5.70,3.07Hz, 1H), 3.22 (br t, J =9.21Hz, 1H), 3.16-3.05 (m, 2H), 2.97 (dd, J=12.06,3.29Hz, 1H), 2.35-2.19 (m, 1H), 1.95 (dt, J=13.37,6.69Hz, 1H).

Embodiment 289

Synthesize (4S)-N- ethyls -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza- 5 (2H)-carboxamide hydrochlorides

To (4S)-N- ethyls -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diazaIn the solution of stirring of -5 (the 2H)-formamides (200mg, 0.531mmol) in 1,4- dioxanes (10mL) The 4M HCl dioxane solution of Isosorbide-5-Nitrae-(5.31mL, 21.26mmol) is added, 3h is then stirred at room temperature.(TLC:Eluant, eluent：5% MeOH/DCM,Rf:0.3).Reactant mixture is concentrated in vacuo and by gained salt with triturated under ether (10mL), obtains desired production Thing (4S)-N- ethyls -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine- 5 (2H)-carboxamide hydrochlorides (130mg, 0.307mmol, 57.7% yield), it is pale solid.LCMS(m/ z):377.07[M+H]⁺, Rt=2.35min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 10.01 (br s, 1H), 8.21 (d, J=7.67Hz, 1H), 8.15 (s, 1H), 7.96 (br d, J=7.89Hz, 1H), 7.85-7.90 (m, 1H), 7.77-7.84 (m, 1H), 7.73 (d, J= 8.11Hz, 1H), 5.53 (dd, J=5.70,3.07Hz, 1H), 3.40-3.64 (m, 4H), 3.27-3.37 (m, 2H), 2.33- 2.46 (m, 1H), 2.02-2.14 (m, 1H), 1.18 (t, J=7.23Hz, 3H).

Embodiment 290

Synthesis (4S)-N- (benzo [d] oxazole -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- bridges Asia Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaNaH (118mg, 2.95mmol) is added in the solution of (300mg, 0.983mmol) in THF (15mL) [d] oxazoles -2- bases-phenyl carbamate (749mg, 2.95mmol), 1h is stirred at 90 DEG C in microwave with benzo.(TLC is eluted Agent：100% ethyl acetate, R_f=0.4；UV activation).Reactant mixture is cooled to room temperature, is quenched with frozen water, is then extracted Into ethyl acetate (2x60mL).The organic extract of merging is filtered and concentrated through anhydrous sodium sulfate drying.Crude compound is passed through Column chromatography purifies (neutral alumina, eluant, eluent：65% ethyl acetate/hexane), obtain desired product (4S)-N- (benzos [d] Oxazole -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (150mg, 0.316mmol, 32.2% yield), it is pale solid.LCMS(m/z):466.10[M +H]⁺,R_t=2.58min.

¹H NMR(400MHz,CDCl₃):δ ppm 14.60 (s, 1H), 8.26 (s, 1H), 8.11 (d, J=7.89Hz, 1H), 7.74-7.81 (m, 1H), 7.72-7.61 (m, 3H), 7.44 (d, J=8.11Hz, 2H), 7.50-7.40 (m, 1H), 7.31- 7.16 (m, 1H), 5.76 (dd, J=6.03,3.18Hz, 1H), 3.34-3.14 (m, 3H), 3.04 (dd, J=12.17, 3.18Hz, 1H), 2.43-2.28 (m, 1H), 2.12 (dt, J=14.20,7.04Hz, 1H).

Embodiment 291

Synthesis (4S)-N- cyclopropyl -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza- 5 (2H)-formamides

Under a nitrogen to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diazaIn the solution of (250mg, 0.819mmol) in THF (20mL) add triethylamine (0.685mL, 4.91mmol), triphosgene (243mg, 0.819mmol), is then stirred at room temperature 30min.Then add cyclopropylamine (140mg, 2.457mmol) and by reactant mixture at 65 DEG C heat 16h.(TLC eluant, eluents：100% ethyl acetate；R_f=0.3；UV is activated ).Removal of solvent under reduced pressure, is diluted with water (20mL) and is extracted with ethyl acetate (2x40mL).The organic extract of merging is through nothing Aqueous sodium persulfate is dried, filtered and concentrated.Crude compound purifies (neutral alumina, eluant, eluent through column chromatography：20% acetic acid second Ester/hexane).The fraction of collection is concentrated under reduced pressure, pure (4S)-N- cyclopropyl -7- (3- (trifluoromethyl) phenyl) -3,4- is obtained Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (170mg, 0.437mmol, 53.4% yield), it is pale solid.LCMS(m/z):389.10[M+H]⁺,R_t=2.34min.

¹H NMR(400MHz,CDCl₃):δ ppm 10.54 (br s, 1H), 8.02 (s, 1H), 7.92 (d, J=7.89Hz, 1H), 7.71-7.65 (m, 1H), 7.64-7.57 (m, 1H), 7.54 (d, J=7.89Hz, 1H), 7.31-7.26 (s, 1H), 5.66 (dd, J=6.03,3.18Hz, 1H), 3.29-3.04 (m, 3H), 2.95 (dd, J=11.84,3.29Hz, 1H), 2.85 (tq, J =7.07,3.62Hz, 1H), 2.27 (dddd, J=14.11,9.95,5.97,3.95Hz, 1H), 2.07-1.96 (m, 1H), 0.84-0.74(m,2H),0.61-0.53(m,2H)。

Embodiment 292

Synthesis (4S)-N- cyclobutyl -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza- 5 (2H)-formamides

Under a nitrogen to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diazaIn the solution of (250mg, 0.819mmol) in THF (20mL) add triethylamine (0.685mL, 4.91mmol), triphosgene (243mg, 0.819mmol), is then stirred at room temperature 30min.Then add cyclobutyl amine (175mg, 2.457mmol) and by reactant mixture at 65 DEG C heat 16h.(TLC eluant, eluents：100% ethyl acetate:R_f=0.3；UV is activated ).Removal of solvent under reduced pressure, is diluted with water (20mL) and is extracted into ethyl acetate (2x40mL).The organic extract warp of merging Anhydrous sodium sulfate drying, filters and concentrates.Crude compound purifies (condition-post through preparation HPLC：XBridge C18 (75X4.6mm,3.5μ)；Mobile phase-A:0.01M ammonium hydrogen carbonate B:Acetonitrile；Gradient-time/%B:0/5,0.8/5,5/50,8/ 95,12/95,12.1/5,15/5；Column temperature：Environment temperature；Flow velocity:1.0ml/min:Diluent：ACN), desired product is obtained (4S)-N- cyclobutyl -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] two Azepine- 5 (2H)-formamides (174mg, 0.432mmol, 52.8% yield), it is pale solid.LCMS(m/z): 403.11[M+H]⁺,R_t=2.56min.

¹H NMR(400MHz,CDCl₃):δ ppm 10.62 (br d, J=6.58Hz, 1H), 8.10 (s, 1H), 7.98 (d, J =7.67Hz, 1H), 7.73-7.65 (m, 1H), 7.64-7.59 (m, 1H), 7.55 (d, J=7.89Hz, 1H), 7.28 (s, 1H), 5.63 (dd, J=6.03,3.18Hz, 1H), 4.43 (dq, J=16.25,7.96Hz, 1H), 3.30-3.02 (m, 3H), 2.94 (dd, J=12.06,3.29Hz, 1H), 2.46-2.34 (m, 2H), 2.25 (dddd, J=14.03,9.98,6.03,3.95Hz, 1H),2.09-1.86(m,3H),1.85-1.63(m,2H)。

Embodiment 293

Synthesize (4S)-N- (6- fluorobenzene simultaneously [d] thiazol-2-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

25 DEG C to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diazaEt is added in the solution of the stirring of (400mg, 1.310mmol) in tetrahydrofuran (15mL)₃N (1.096mL, 7.86mmol) and triphosgene (389mg, 1.310mmol) and stirring 1h, then add 6- fluorobenzene simultaneously [d] thiazole- 2- amine (441mg, 2.62mmol) (solid) simultaneously heats 15h at 70 DEG C.(TLC systems:Net ethyl acetate, Rf:0.3).Will reaction Mixture is cooled to room temperature, is concentrated in vacuo and by residue distribution between water (40mL) and EtOAc (80mL).Separate organic layer And through anhydrous sodium sulfate drying, filter and filtrate evaporation is obtained into crude compound.Crude product is through flash column chromatography (100- 200 silica gel, eluant, eluent：80% ethyl acetate/hexane), obtain desired product (4S)-N- (6- fluorobenzene simultaneously [d] thiazole -2- Base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (170mg, 0.333mmol, 25.4% yield), it is pale solid.LCMS(m/z):500.01[M+H ]⁺, Rt=3.10min.

¹H NMR(400MHz,DMSO-d₆):δppm 14.75(s,1H),8.42(s,2H),7.94-7.75(m,5H), 7.68 (dd, J=8.77,4.82Hz, 1H), 7.31 (td, J=9.10,2.85Hz, 1H), 5.50 (dd, J=6.03,3.18Hz, 1H), 3.27-3.09 (m, 3H), 2.99 (dd, J=11.73,3.18Hz, 1H), 2.35-2.22 (m, 1H), 2.10-1.96 (m, 1H)。

Embodiment 294

Synthesize (4S) -7- (2- picoline -4- bases)-N- (4- (3- (trifluoromethyl) phenyl) pyridine -2- bases) -3,4- two Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4S)-N- (4- bromopyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (300mg, 0.665mmol), (3- (trifluoromethyl) phenyl) boric acid (189mg, 0.997mmol) and tripotassium phosphate (423mg, 1.994mmol) are in 1,4- dioxanes (18mL) and water (4.50mL) X-phos (19.20mg, 0.133mmol) and Pd is added in the solution of degassing₂(dba)₃(60.9mg,0.066mmol).Will reaction Mixture stirs 16h at 100 DEG C.(TLC systems:5%MeOH/EtOAc, R_f:0.5).So that reactant mixture is cooled to room temperature And 1,4 dioxane solvents are evaporated under reduced pressure, by the residue diluted with water (50ml) of acquisition and it is extracted with ethyl acetate (2x100ml).The organic layer water of merging, salt water washing, it is dried over sodium sulfate and solvent is evaporated under reduced pressure, obtain crude product.Slightly Product uses (silica gel 100-200 mesh through flash column chromatography：Eluant, eluent：Net ethyl acetate), obtain desired product (4S) -7- (2- picoline -4- bases)-N- (4- (3- (trifluoromethyl) phenyl) pyridine -2- bases) -3,4- dihydro -1,4- bridges are sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (70mg, 0.135mmol, 20.34% yield), it is Pale solid.LCMS(m/z):517.11[M+H]⁺,R_t=2.16min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.70 (s, 1H), 8.64 (d, J=5.26Hz, 1H), 8.57-8.53 (m, 1H), 8.45 (d, J=5.04Hz, 1H), 8.22 (s, 1H), 7.85-7.99 (m, 2H), 7.76-7.57 (m, 4H), 7.50 (d, J=7.89Hz, 1H), 7.27-7.20 (m, 1H), 5.72 (dd, J=5.92,3.07Hz, 1H), 3.34-3.13 (m, 3H), 3.04 (dd, J=12.06,3.29Hz, 1H), 2.77 (s, 3H), 2.46-2.29 (m, 1H), 2.21-2.02 (m, 1H).

Embodiment 295

(4S)-N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (2- picoline -4- bases) -3 is synthesized, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamide

0 DEG C to (4S)-N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine - 4- yls) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 In the solution of stirring of (the 2H)-formamide (150mg, 0.298mmol) in methanol (10mL) add hydrochloric acid (5mL, 165mmol), 5min a period of time is lasted.Then reactant mixture is stirred into 2h at 30 DEG C.(TLC eluant, eluents：5%MeOH/ DCM:R_f-0.5；UV activation), then reactant mixture is neutralized, is extracted with DCM, so by solvent evaporation with sodium bicarbonate solution Solid chemical compound is obtained by anhydrous sodium sulfate drying and evaporation, filtering is simultaneously washed (2 × 20mL) with pentane, obtains desired Product (4S)-N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (2- picoline -4- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(110mg, 0.238mmol, 80% are received -5 (2H)-formamides Rate), it is pale solid.LCMS(m/z):463.15[M+H]⁺,R_t=1.20min.

¹H NMR(400MHz,CDCl3):δ ppm 13.28 (s, 1H), 8.68 (d, J=5.26Hz, 1H), 7.96 (d, J= 5.70Hz, 1H), 7.65 (d, J=7.89Hz, 1H), 7.58 (s, 1H), 7.48 (dd, J=5.15,1.21Hz, 1H), 7.38 (d, J=7.89Hz, 1H), 7.13-7.05 (m, 2H), 5.67 (dd, J=6.03,3.18Hz, 1H), 4.46 (dd, J=4.71, 1.21Hz, 2H), 4.35 (d, J=5.48Hz, 1H), 3.99 (dq, J=9.95,4.94Hz, 1H), 3.73-3.61 (m, 2H), 3.33-3.12 (m, 3H), 3.03 (dd, J=12.17,3.18Hz, 1H), 2.89 (t, J=6.58Hz, 1H), 2.69 (s, 3H), 2.35 (qd, J=9.94,4.17Hz, 1H), 2.08 (dt, J=14.14,6.96Hz, 1H).

Embodiment 296

Synthesize (4S) -7- (2- picoline -4- bases)-N- (4- (2- methylthiazol -5- bases) pyridine -2- bases) -3,4- two Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4S)-N- (4- bromopyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (250mg, 0.554mmol), 2- methyl -5- (4,4,5,5- tetramethyls Base -1,3,2- dioxaborolan -2- bases) thiazole (249mg, 1.108mmol) and tripotassium phosphate (353mg, 1.662mmol) In 1,4- dioxanes (18mL) and water (4.50mL) PdCl is added in the solution of degassing₂(dppf)-CH₂Cl₂Adduct (45.2mg,0.055mmol).Reactant mixture is stirred into 16h at 90 DEG C.(TLC:SiO2；10%MeOH/DCM TLC:SiO2； 10%MeOH/DCM).So that reactant mixture is cooled to room temperature and is diluted with water (50mL), it is extracted with ethyl acetate (2x50mL).(50mL) is washed with water in the organic layer of merging, through anhydrous sodium sulfate drying and is evaporated in vacuo, and obtains crude product, is Brown solid.(120g is anti-phase by combi purification by flash chromatography for crude product:100% methanol), obtain desired product (4S)- 7- (2- picoline -4- bases)-N- (4- (2- methylthiazol -5- bases) pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (67mg, 0.142mmol, 25.6% yield), it is solid for light green color Body.LCMS(m/z):470.09[M+H]⁺,R_t=1.63min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.65 (s, 1H), 8.63 (d, J=5.26Hz, 1H), 8.45 (dd, J= 1.53,0.66Hz, 1H), 8.35 (dd, J=5.15,0.77Hz, 1H), 8.18 (d, J=1.53Hz, 1H), 8.05 (s, 1H), 7.72 (dd, J=5.37,1.43Hz, 1H), 7.63 (d, J=7.89Hz, 1H), 7.49 (d, J=7.89Hz, 1H), 7.14 (dd, J=5.26,1.75Hz, 1H), 5.72 (dd, J=6.03,3.18Hz, 1H), 3.40-3.14 (m, 3H), 3.03 (dd, J= 12.06,3.29Hz, 1H), 2.75 (d, J=6.58Hz, 6H), 2.36 (dddd, J=14.11,9.95,6.08,4.06Hz, 1H), 2.10 (dt, J=14.25,6.91Hz, 1H).

Embodiment 297

Synthesize (4S)-N- (4- methoxyl groups benzo [d] isoxazole -3-base) -7- (3- (trifluoromethyl) phenyl) -3,4- two Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

25 DEG C to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diazaThree second are added in the solution of the stirring of (350.0mg, 1.146mmol) in tetrahydrofuran (20mL) Amine (0.799mL, 5.73mmol) and triphosgene (340mg, 1.146mmol) and stirring 45min, then add 4- methoxyl group benzos [d] isoxazole -3- amine (565mg, 3.44mmol).Reactant mixture is heated into 6h at 72 DEG C.(TLC eluent systems:100% EtOAc, Rf-0.4, UV activation).Reactant mixture is cooled to room temperature, the solid of precipitation is filtered and filtrate water is washed Wash (10mL) and be extracted into ETOAc.Separate organic layer and through anhydrous sodium sulfate drying, filter and filtrate evaporation is obtained rough Compound.Crude product purifies (condition through preparation HPLC:Post XBridge C18 (250X30mm, 5 μ)；Mobile phase-A:5mM carbon Sour hydrogen ammonium B:Acetonitrile；Gradient-time/%B:0/10,10/50,11.5/100,15/100,15.1/10；Column temperature：Environment temperature Degree；Flow velocity:20ml/min:Diluent：THF+MEOH+ACN), required product (4S)-N- (4- methoxyl groups benzo [d] different Evil are obtained Azoles -3- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza - 5 (2H)-formamides (106.5mg, 0.211mmol, 18.38% yield), it is white solid LCMS (m/z):496.05[M+ H]⁺,R_t=2.43min.

¹HNMR(400MHz,CDCl₃):δ ppm 12.97 (s, 1H), 8.06 (s, 1H), 7.99 (d, J=7.89Hz, 1H), 7.62-7.68 (m, 2H), 7.45 (t, J=7.78Hz, 1H), 7.35-7.41 (m, 2H), 7.10 (d, J=8.11Hz, 1H), 6.34 (d, J=7.89Hz, 1H), 5.77 (dd, J=5.81,3.18Hz, 1H), 3.18-3.35 (m, 3H), 3.03 (dd, J= 12.06,3.29Hz, 1H), 2.90 (s, 3H), 2.30-2.41 (m, 1H), 2.16 (dt, J=14.03,7.02Hz, 1H)

Embodiment 298

Synthesize (4S)-N- (1H- benzos [d] imidazoles -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides:

Room temperature to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diazaTEA is added in the solution of the stirring of (300mg, 0.983mmol) in tetrahydrofuran (40mL) (0.685mL, 4.91mmol) and triphosgene (292mg, 0.983mmol) and stirring 30min.Then 1H- benzos [d] miaow is added Reactant mixture is simultaneously heated to 60 DEG C by azoles -2- amine (393mg, 2.95mmol), keeps 16h.Solid body (TLC elution systems: 10%MeOH/DCM；R_f-0.3；UV activation).Reactant mixture is cooled to room temperature and (15mL) is quenched with water and is extracted into In EtOAc (2x15mL).Organic layer is separated, through anhydrous sodium sulfate drying, filters and filtrate evaporation is obtained into crude compound.Will Through chromatogram purification, (GRACE uses C-18 reversed-phase columns, mobile phase A to thick material:0.1% formic acid/water；B:MeOH, eluant, eluent 75%B/ A).The fraction of merging is evaporated, saturation NaHCO is then used₃It is basified.Water layer is extracted with DCM, DCM layers through anhydrous Na₂SO₄ Dry, filter simultaneously filtrate is evaporated, obtain (4S)-N- (1H- benzos [d] imidazoles -2- bases) -7- (3- (trifluoromethyl) phenyl) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (26mg, 0.053mmol, 5.43% yield), it is canescence LCMS (m/z):465.05[M+H]⁺,R_t=2.32min.

¹H NMR(400MHz,DMSO-d₆):δppm 14.16(s,1H),11.99(s,1H),8.40-8.50(m,2H), 7.86-7.92 (m, 1H), 7.79-7.86 (m, 2H), 7.74-7.78 (m, 1H), 7.46-7.51 (m, 1H), 7.38 (d, J= 7.45Hz, 1H), 7.08 (td, J=7.73,1.43Hz, 2H), 5.53 (dd, J=5.81,2.96Hz, 1H), 3.09-3.27 (m, 3H), 3.00 (dd, J=11.95,3.40Hz, 1H), 2.23-2.35 (m, 1H), 2.03 (dt, J=13.98,7.15Hz, 1H)

Embodiment 299

Synthesize (4S)-N- (4- (1,3- dimethyl -1H- pyrazoles -4- bases) pyridine -2- bases) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4S)-N- (4- bromopyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (250mg, 0.554mmol), 1,3- dimethyl -4- (4,4,5,5- Tetramethyl -1,3,2- dioxaborolan -2- bases) -1H- pyrazoles (135mg, 0.609mmol) and sodium carbonate (176mg, 1.662mmol) at 1,4- dioxanes (7mL):Pd (P (Ph) are added in water (3mL) in the solution of degassing₃)₄(32.0mg, 0.028mmol) and by reactant mixture at 80 DEG C stir 15h.(TLC systems:5% methanol/ethyl acetate .Rf values：0.3.).So After allow to cool to room temperature and be diluted with water (30mL), be extracted with ethyl acetate (2X50mL).The organic layer of merging is water-soluble with salt Liquid washs (20mL), through anhydrous sodium sulfate drying, filters and concentrates, obtain crude compound.Crude compound is through quick post color Spectrum purifying (silica gel:100-200 mesh, eluant, eluent：2%MeOH/ ethyl acetate), obtain desired product (4S)-N- (4- (1,3- Dimethyl -1H- pyrazoles -4- bases) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides (150mg, 0.321mmol, 57.9% yield), it is solid for canescence Body.LCMS(m/z):467.18[M+H]⁺,R_t=1.47min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.56 (s, 1H), 8.62 (d, J=5.26Hz, 1H), 8.37-8.28 (m, 2H), 8.23 (s, 1H), 7.73 (d, J=1.32Hz, 1H), 7.67 (s, 1H), 7.63 (s, 1H), 7.49 (d, J= 7.89Hz, 1H), 7.09-7.04 (m, 1H), 5.71 (dd, J=5.81,3.18Hz, 1H), 3.89 (s, 3H), 3.32-3.15 (m, 3H), 3.02 (dd, J=12.06,3.29Hz, 1H), 2.75 (s, 3H), 2.52 (s, 3H), 2.38-2.30 (m, 1H), 2.15- 2.05(m,1H)。

Embodiment 300

Synthesize (4S) -7- (6- aminopyridine -3- bases)-N- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the chloro- N- of (4S) -7- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (350mg, 1.108mmol), (6- aminopyridine -3- bases) boric acid (229mg, 1.663mmol) and phosphorus Sour tripotassium (235mg, 1.108mmol) is at 1,4- dioxanes (18mL):In water (4.50mL) Pd is added in the solution of degassing₂ (dba)₃(50.8mg, 0.055mmol) and x-phos (52.8mg, 0.111mmol).Reactant mixture is stirred into 3h at 100 DEG C. (TLC:10%MeOH/EtOAc Rf:0.3).1,4 dioxane solvents are evaporated under reduced pressure, by the residue diluted with water of acquisition (50mL) and it is extracted with ethyl acetate (2x100ml).The organic layer water of merging, salt water washing, it is dried over sodium sulfate and depressurize Evaporation solvent, obtains crude product.Crude product is through flash column chromatography ((silica gel 100-200 mesh：Eluant, eluent：2%MeOH/ DCM), obtain half pure compound and purify (preparation HPLC condition again by preparation HPLC：MP-A5mM ammonium acetate (water Solution)) MP-B:MeOH+ACN posts:XBRIDGE250X30 methods：- T/%B=0/20,10/50,11/100 flow velocitys：28ml/ Min eluant, eluents：THF+MeOH), desired product (4S) -7- (6- aminopyridine -3- bases)-N- (pyridine -2- bases) -3,4- are obtained Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (210mg, 0.562mmol, 50.7% yield), it is pale solid.LCMS(m/z):[374.19 M+H]+, Rt=1.32min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.64 (s, 1H), 8.75 (d, J=2.41Hz, 1H), 8.50 (dd, J= 8.77,2.41Hz, 1H), 8.38-8.34 (m, 1H), 8.18 (dt, J=8.55,0.88Hz, 1H), 7.68 (td, J=7.89, 1.97Hz, 1H), 7.54 (d, J=8.11Hz, 1H), 7.29 (d, J=8.11Hz, 1H), 7.02-6.94 (m, 1H), 6.67 (dd, J=8.55,0.66Hz, 1H), 5.68 (dd, J=5.92,3.29Hz, 1H), 4.65 (br s, 2H), 3.32-3.12 (m, 3H), 2.99 (dd, J=12.06,3.29Hz, 1H), 2.31 (dddd, J=13.95,9.95,6.03,3.84Hz, 1H), 2.12-2.03 (m,1H)。

Embodiment 301

(4S)-N- (4- (1- methyl isophthalic acid H- imidazoles -5- bases) pyridine -2- bases) -7- (2- picoline -4- bases) -3 is synthesized, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To 1- methyl -5- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) -1H-imidazole (138mg, 0.665mmol) is at 1,4- dioxanes (8mL):Added in the solution of degassing in water (2mL) (4S)-N- (4- bromopyridines- 2- yls) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (200mg, 0.443mmol), Na₂CO₃(47.0mg,0.443mmol).Then by Pd (Ph₃P)₄(512mg, Reactant mixture 0.443mmol) is added in room temperature and the 5min that deaerates again.Then reactant mixture is stirred into 16h at 80 DEG C. (TLC systems:(10%MeOH/DCM, Rf:0.5) reactant mixture, is cooled to room temperature, organic solvent evaporation is used in combination Water dilutes, and is then extracted with ethyl acetate (3X20mL).The organic layer of merging is with salt water washing (2x5mL), through anhydrous sodium sulfate It is dried, filtered and concentrated, obtains crude compound.Crude compound is through flash column chromatography (100-200 mesh, net acetic acid second Ester -2%DCM/MeOH), obtain desired product (4S)-N- (4- (1- methyl isophthalic acid H- imidazoles -5- bases) pyridine -2- bases) -7- (2- Picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (70mg, 0.154mmol, 34.7% yield), it is yellow solid.LCMS(m/z):453.14 [M+H]+, Rt=1.18min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.63 (s, 1H), 8.60 (d, J=5.26Hz, 1H), 8.42 (d, J =5.26Hz, 1H), 8.33 (s, 1H), 8.23 (s, 1H), 7.94 (d, J=4.60Hz, 1H), 7.86-7.77 (m, 2H), 7.75- 7.68 (m, 1H), 7.39-7.27 (m, 2H), 5.52 (br d, J=2.41Hz, 1H), 3.83 (s, 3H), 3.45-2.93 (m, 4H),2.64(s,3H),2.35-2.15(m,1H),2.09-1.86(m,1H)。

Embodiment 302

Synthesize (4S) -7- (6- (dimethylamino) -5- fluorine pyridin-3-yl)-N- (pyridine -2- bases) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the chloro- N- of (4S) -7- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (3.2g, 10.13mmol), (6- (dimethylamino) -5- fluorine pyridin-3-yl) boric acid (2.051g, 11.15mmol) with sodium carbonate (3.22g, 30.4mmol) at 1,4- dioxanes (18mL):In water (4.50mL) in the solution of degassing Add Pd (PPh₃)₄(0.586g, 0.507mmol) and stir 3h at 100 DEG C.(TLC systems:Net ethyl acetate, Rf:0.4) subtracts Pressure evaporation 1,4- dioxane solvents, by the residue diluted with water (50mL) of acquisition and are extracted with ethyl acetate (2x100ml).Close And organic layer water, salt water washing, it is dried over sodium sulfate and solvent is evaporated under reduced pressure, obtain crude product.Crude product is through quick post Chromatogram purification (silica gel 100-200 mesh：Eluant, eluent：70% ethyl acetate), obtain desired product (4S) -7- (6- (dimethylaminos Base) -5- fluorine pyridin-3-yl)-N- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (1.58g, 3.76mmol, 37.1% yield), it is pale solid.LCMS(m/z):420.1[M +H]⁺, Rt=2.25min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.76 (s, 1H), 8.60 (d, J=1.97Hz, 1H), 8.56 (s, 1H), 8.38 (dd, J=4.82,1.10Hz, 1H), 8.16 (d, J=8.55Hz, 1H), 7.77-7.64 (m, 1H), 7.53 (d, J= 8.11Hz, 1H), 7.31 (d, J=8.11Hz, 1H), 7.02-6.91 (m, 1H), 5.67 (dd, J=5.92,3.07Hz, 1H), 3.25 (s, 3H), 3.23 (s, 3H), 3.22-3.02 (m, 3H), 2.98 (dd, J=11.84,3.29Hz, 1H), 2.31 (dddd, J =14.06,9.95,6.03,3.95Hz, 1H), 2.07 (dt, J=13.98,6.93Hz, 1H).

Embodiment 303

Synthesis (4S)-N- (2- (3- hydroxy propyloxy groups) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diazaTriphosgene is added in the solution of the stirring of (300mg, 0.983mmol) in THF (15mL) (175mg, 0.590mmol) and triethylamine (0.822mL, 5.90mmol) and stirring 30min.Then, ((4- amino is phonetic by addition 3- Pyridine -2- bases) epoxide) propyl- 1- alcohol (332mg, 1.965mmol), then stirs 15h at 80 DEG C.(TLC systems:R_f- 0.2,5% MeOH/EtOAc).So that reactant mixture is cooled to room temperature and is diluted with water (25mL), extracted with EtOAc (2x40mL).Merge Organic layer through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain crude compound.Crude compound is through quick post color Spectrum purifying (silica gel：100-200 mesh, eluant, eluent：2% methanol/ethyl acetate), obtain desired product (4S)-N- (2- (3- hydroxyls Base propoxyl group) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (80mg, 0.152mmol, 15.46% yield), it is pale solid.LCMS(m/ z):501.24[M+H]⁺,R_t=2.19min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.65 (s, 1H), 8.46 (dd, J=4.17,2.85Hz, 1H), 8.36 (d, J=5.70Hz, 1H), 8.06 (s, 1H), 7.79 (d, J=5.48Hz, 1H), 7.73 (s, 2H), 7.65 (d, J=8.11Hz, 1H), 7.43 (d, J=7.89Hz, 1H), 5.66 (dd, J=5.92,3.07Hz, 1H), 4.45-4.40 (m, 2H), 3.76 (q, J =5.19Hz, 2H), 3.36-3.14 (m, 3H), 3.02 (dd, J=12.06,3.29Hz, 1H), 2.50-2.28 (m, 1H), 2.14-1.88(m,4H)。

Embodiment 304

Synthesize (4S)-N- (6- (3- hydroxyl -2,2- dimethyl propylenes epoxide) pyrazine -2- bases) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diazaTriphosgene is added in the solution of the stirring of (300mg, 0.983mmol) in THF (15mL) (175mg, 0.590mmol) and triethylamine (0.822mL, 5.90mmol) and stirring 30min.Then, 3- ((6- amino pyrroles are added Piperazine -2- bases) epoxide) -2,2- dimethyl propylene -1- alcohol (388mg, 1.965mmol), then stirs 15h at 80 DEG C.(TLC systems: R_f- 0.4,5%MeOH/EtOAc).So that reactant mixture is cooled to room temperature and is diluted with water (25mL), extracted with EtOAc (2x40mL).The organic layer of merging is through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain crude compound.Rough chemical combination Thing is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：1% methanol/ethyl acetate), obtain desired product (4S)-N- (6- (3- hydroxyl -2,2- dimethyl propylenes epoxide) pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(130mg, 0.240mmol, 24.40% are received -5 (2H)-formamides Rate), it is pale solid.LCMS(m/z):529.17[M+H]⁺,R_t=2.52min.

¹H NMR(400MHz,CDCl₃):δppm 13.24(s,1H),8.99(s,1H),8.24-8.18(m,1H),8.05 (s, 1H), 7.94 (s, 1H), 7.71-7.66 (m, 2H), 7.64 (d, J=7.89Hz, 1H), 7.38 (d, J=7.89Hz, 1H), 5.71 (dd, J=5.92,3.07Hz, 1H), 3.89 (s, 2H), 3.35-3.13 (m, 5H), 3.02 (dd, J=12.06, 3.29Hz, 1H), 2.40-2.29 (m, 1H), 2.14-1.99 (m, 2H), 0.88 (d, J=2.41Hz, 6H).

Embodiment 305

(4S)-N- (2- (3- hydroxy-3-methyls butoxy) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3 is synthesized, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diazaTriphosgene is added in the solution of the stirring of (300mg, 0.983mmol) in THF (15mL) (175mg, 0.590mmol) and triethylamine (0.822mL, 5.90mmol), and stirring 30min.Then, ((4- amino is phonetic by addition 4- Pyridine -2- bases) epoxide) -2- methyl butyl- 2- alcohol (388mg, 1.965mmol), then stirs 15h at 80 DEG C.(TLC systems:R_f- 0.3,5%MeOH/EtOAc).So that reactant mixture is cooled to room temperature and is diluted with water (25mL), extracted with EtOAc (2x40mL).The organic layer of merging is through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain crude compound.Rough chemical combination Thing is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：1% methanol/ethyl acetate), obtain desired product (4S)-N- (2- (3- hydroxy-3-methyls butoxy) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (115mg, 0.217mmol, 22.04% yield), It is pale solid.LCMS(m/z):529.25[M+H]⁺,R_t=2.37min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.62 (s, 1H), 8.54-8.46 (m, 1H), 8.37 (d, J= 5.70Hz, 1H), 8.04 (s, 1H), 7.80 (d, J=5.70Hz, 1H), 7.75-7.69 (m, 2H), 7.64 (d, J=8.11Hz, 1H), 7.43 (d, J=8.11Hz, 1H), 5.66 (dd, J=5.92,3.07Hz, 1H), 4.49-4.39 (m, 2H), 3.37-3.14 (m, 3H), 3.02 (dd, J=12.06,3.29Hz, 1H), 2.40-2.29 (m, 1H), 2.17-2.01 (m, 2H), 1.97 (t, J= 6.47Hz,2H),1.28(s,6H)。

Embodiment 306

(4S)-N- (4- (1- methyl isophthalic acid H- pyrazole-3-yls) pyridine -2- bases) -7- (2- picoline -4- bases) -3 is synthesized, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

It is sub- to (4S)-N- (4- bromopyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridges in room temperature Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (200mg, 0.443mmol), 1- methyl -3- (4,4,5, 5- tetramethyl -1,3,2- dioxaborolan -2- bases) -1H- pyrazoles (138mg, 0.665mmol) and K₃PO₄(282mg, 1.329mmol) at 1,4- dioxanes (8mL):In water (2mL) PdCl is added in the solution of degassing₂(dppf)(64.9mg, 0.089mmol) and by reactant mixture at 100 DEG C stir 6h.(TLC systems:5% ethanol/methylene, R_f:0.2).Will be anti- Mixture is answered to pour into cold water (10mL) and be extracted with ethyl acetate (2x50mL).The organic layer of merging is dry through anhydrous sodium sulfate It is dry, filter and be concentrated under reduced pressure, obtain crude compound.Crude compound is through flash column chromatography (neutral alumina, 2% first Alcohol/DCM), obtain desired product (4S)-N- (4- (1- methyl isophthalic acid H- pyrazole-3-yls) pyridine -2- bases) -7- (2- methyl pyrroles Pyridine -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (110mg, 0.241mmol, 54.3% yield), it is pale solid.LCMS(m/z):453.14[M+H]⁺,R_t=1.49min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.56 (s, 1H), 8.67-8.58 (m, 2H), 8.37 (d, J= 5.26Hz, 1H), 8.23 (s, 1H), 7.72 (dd, J=5.15,1.43Hz, 1H), 7.62 (d, J=7.89Hz, 1H), 7.55- 7.46 (m, 2H), 7.41 (d, J=2.19Hz, 1H), 6.74 (d, J=2.19Hz, 1H), 5.73 (dd, J=5.92,3.29Hz, 1H), 3.98 (s, 3H), 3.14-3.37 (m, 3H), 3.03 (dd, J=12.06,3.29Hz, 1H), 2.76 (s, 3H), 2.41- 2.29(m,1H),2.19-2.03(m,1H)。

Embodiment 307

Synthesize (4S)-N- (4- (1- methyl -3- (trifluoromethyl) -1H- pyrazoles -4- bases) pyridine -2- bases) -7- (2- methyl Pyridin-4-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

It is sub- to (4S)-N- (4- bromopyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridges in room temperature Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (200mg, 0.443mmol), 1- methyl -4- (4,4,5, 5- tetramethyl -1,3,2- dioxaborolan -2- bases) -3- (trifluoromethyl) -1H- pyrazoles (184mg, 0.665mmol) and K₃PO₄(282mg, 1.329mmol) is at 1,4- dioxanes (8mL):In water (2mL) PdCl is added in the solution of degassing₂(dppf) (32.4mg, 0.044mmol) and reactant mixture is stirred into 6h at 100 DEG C.(TLC systems:Net ethyl acetate, Rf:0.3).Will Reactant mixture is poured into cold water (10mL) and is extracted with ethyl acetate (2x50mL).The organic layer of merging is dry through anhydrous sodium sulfate It is dry, filter and be concentrated under reduced pressure, obtain crude compound.Crude compound is through flash column chromatography (silica gel:100-200 mesh, is washed De- agent：2% methanol/DCM), obtain desired product (4S)-N- (4- (1- methyl -3- (trifluoromethyl) -1H- pyrazoles -4- bases) Pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (60mg, 0.115mmol, 25.9% yield), it is faint yellow solid.LCMS(m/z):521.14[M+ H]⁺, Rt=1.75min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.64 (s, 1H), 8.63 (d, J=5.48Hz, 1H), 8.38 (d, J= 5.04Hz, 1H), 8.32 (s, 1H), 8.21 (s, 1H), 7.77-7.70 (m, 2H), 7.63 (d, J=7.89Hz, 1H), 7.49 (d, J=7.89Hz, 1H), 7.12 (d, J=5.04Hz, 1H), 5.70 (dd, J=5.92,3.07Hz, 1H), 4.00 (s, 3H), 3.37-3.14 (m, 3H), 3.02 (dd, J=12.28,3.29Hz, 1H), 2.76 (s, 3H), 2.41-2.26 (m, 1H), 2.17- 2.02(m,1H)。

Embodiment 308

Synthesize (4S)-N- (pyridine -2- bases) -7- ((R) -4- (trifluoromethyl) -4,5- dihydro-oxazole -2- bases) -3,4- two Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides:

Under a nitrogen in room temperature to fluoro- 2- ((4S) -5- (pyridine -2- base carbamyls of methanesulfonic acid (2R) -3,3,3- three Base) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 7- formamido groups) propyl diester Sodium azide (190mg, 2.92mmol) is added in the solution of (300mg, 0.583mmol) in DMSO (5mL) and is stirred at 90 DEG C Mix 1h.(TLC systems:5% methanol/DCM.Rf values：0.5).Reactant mixture frozen water is diluted into (25ml), EtOAc is then used (2x50mL) is extracted.The organic layer of merging is dried over sodium sulfate with salt water washing (25mL), filters and is concentrated under reduced pressure, and obtains thick Produced compounds.Thick material through combiflash chromatogram purifications (using silicagel column, 5%MeOH/DCM), obtain (4S)-N- (pyridine- 2- yls) -7- ((R) -4- (trifluoromethyl) -4,5- dihydro-oxazole -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza- 5 (2H)-formamides (70mg, 0.164mmol, 28.1% yield), it is pale solid.LC-MS (m/z):419.04[M+H]⁺, Rt=1.61min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.03 (s, 1H), 8.34 (dd, J=4.82,1.10Hz, 1H), 8.08 (d, J=8.55Hz, 1H), 7.56-7.77 (m, 3H), 6.98 (ddd, J=7.34,4.93,0.88Hz, 1H), 5.69 (dd, J= 6.03,3.18Hz, 1H), 4.90 (dquin, J=10.02,7.31,7.31,7.31,7.31Hz, 1H), 4.59-4.78 (m, 2H),3.08-3.30(m,3H),2.92-3.06(m,1H),2.19-2.39(m,1H),1.93-2.17(m,1H)。

Embodiment 309

(4S)-N- (6- (3- hydroxy-3-methyls butoxy) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3 is synthesized, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides:

Room temperature to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diazaIn the solution of the stirring of (500mg, 1.638mmol) in THF (25mL) add TEA (1.141mL, 8.19mmol), triphosgene (292mg, 0.983mmol), stirs 30min, then adds 4- ((6- aminopyridine -2- bases) oxygen Base) -2- methyl butyl- 2- alcohol (643mg, 3.28mmol) and by reactant mixture 100 DEG C stir 16h.(TLC eluent systems: 100%EtOAc, Rf-0.4, UV activation).Reactant mixture is cooled to room temperature, then distributed in water (25mL) and EtOAc Between (2x25mL).Organic layer is separated, through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into crude product.Crude compound Through chromatogram purification (neutral alumina, eluant, eluent：20% ethyl acetate/hexane), obtain (4S)-N- (6- (3- hydroxy-3-methyls Butoxy) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (120mg, 0.225mmol, 13.72% yield), it is pale solid LCMS (m/z): 528.20[M+H]⁺,R_t=2.64min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.97 (s, 1H), 8.31 (br d, J=7.23Hz, 1H), 8.04 (s, 1H), 7.73 (d, J=7.89Hz, 1H), 7.55-7.70 (m, 4H), 7.36 (d, J=7.89Hz, 1H), 6.42 (d, J= 8.11Hz, 1H), 5.70 (dd, J=5.70,3.07Hz, 1H), 4.18 (td, J=6.14,1.97Hz, 2H), 3.11-3.37 (m, 3H), 3.01 (dd, J=12.06,3.07Hz, 1H), 2.25-2.40 (m, 1H), 2.03-2.15 (m, 2H), 1.81 (t, J= 6.25Hz, 2H), 1.18 (d, J=1.32Hz, 6H).

Embodiment 310

Synthesize (4R) -7- (2- picoline -4- bases)-N- (4- (oxazole -5- bases) pyridine -2- bases) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4R)-N- (4- bromopyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (400mg, 0.886mmol), K₃PO₄(564mg,2.66mmol) (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- base) oxazoles (259mg, 1.329mmol) are in 1,4- bis- Evil with 5- In alkane (12mL) and water (3mL) PdCl is added in the solution of degassing₂(dppf)-CH₂Cl₂(36.2mg,0.044mmol).Will reaction Mixture deaerates 5min again.Then reactant mixture is stirred into 16h at 80 DEG C.(TLC systems:(10%MeOH/DCM, Rf: 0.4) room temperature, is then allowed to cool to, is diluted with water and is extracted with ethyl acetate (3X50mL).The organic layer of merging is washed with salt (2x5mL) is washed, through anhydrous sodium sulfate drying, filters and concentrates, crude residue is obtained.Crude compound is pure through flash column chromatography Change (100-200 mesh, net ethyl acetate -2%DCM/MeOH), obtain required product (4R) -7- (2- picoline -4- bases)-N- (4- (oxazole -5- bases) pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (270mg, 0.613mmol, 69.2% yield), it is yellow solid.LCMS(m/z):440.18[M+H]⁺, Rt:1.46min。

¹H NMR(400MHz,CDCl₃):δ ppm 13.69 (s, 1H), 8.63 (d, J=5.48Hz, 1H), 8.56 (s, 1H), 8.42 (d, J=5.04Hz, 1H), 8.18 (s, 1H), 7.99 (s, 1H), 7.72 (dd, J=5.15,1.43Hz, 1H), 7.66- 7.61 (m, 2H), 7.50 (d, J=8.11Hz, 1H), 7.27 (s, 1H), 5.72 (dd, J=5.92,3.07Hz, 1H), 3.37- 3.15 (m, 3H), 3.04 (dd, J=12.17,3.40Hz, 1H), 2.75 (s, 3H), 2.44-2.30 (m, 1H), 2.19-2.06 (m,1H)。

Embodiment 311

Synthesis (4S)-N- (4- (2,2- dimethyl tetrahydro -2H- pyrans -4- bases) pyridine -2- bases) -7- (2- picolines - 4- yls) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (4S)-N- (4- (2,2- dimethyl -3,6- dihydro -2H- pyrans -4- bases) pyridine -2- bases) -7- (2- Picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides 10% is added in the solution of the stirring of (220mg, 0.456mmol) and ammonium formate (287mg, 4.56mmol) in methanol (30mL) Pd-C (48.5mg, 0.046mmol) and 65 DEG C stir 16h.(TLC eluant, eluents：5%MeOH/DCM:R_f-0.3；UV is activated ).Reactant mixture is cooled down, solid filters through diatomite and concentrates filtrate, obtains crude product.By crude product pentane (5mL) and ether (5ml) are ground, and obtain desired (4S)-N- (4- (2,2- dimethyl tetrahydro -2H- pyrans -4- bases) pyridine -2- Base) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (57mg, 0.117mmol, 25.7% yield), it is pale solid.LCMS(m/z):485.25[M+H]⁺, R_t=1.67min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.54 (s, 1H), 8.61 (d, J=5.26Hz, 1H), 8.29 (d, J= 5.26Hz, 1H), 8.20 (s, 1H), 8.14 (s, 1H), 7.71 (dd, J=5.26,1.53Hz, 1H), 7.62 (d, J=7.89Hz, 1H), 7.48 (d, J=8.11Hz, 1H), 6.89 (dd, J=5.04,1.32Hz, 1H), 5.71 (dd, J=5.81,3.18Hz, 1H),3.90-3.74(m,2H),3.36-3.14(m,3H),3.06-2.92(m,2H),2.74(s,3H),2.40-2.30(m, 1H), 2.09 (dt, J=14.14,6.96Hz, 1H), 1.84-1.63 (m, 3H), 1.59 (s, 1H), 1.30 (d, J=13.81Hz, 6H)

Embodiment 312

Synthesis (4S)-N- (6- (3- hydroxy propyloxy groups) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Under a nitrogen in room temperature to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diazaEt is added in the solution of the stirring of (0.4g, 1.310mmol) in THF (50mL)₃N (0.913mL, 6.55mmol), triphosgene (0.389g, 1.310mmol) simultaneously stir 1h.Then 3- ((6- aminopyridine -2- bases) Epoxide) propyl- 1- alcohol (0.331g, 1.965mmol) add and by reactant mixture 65 DEG C heat 16h.(TLC eluant, eluents： 100%EtOAc:R_f-0.3；UV activation).Reactant mixture is cooled to room temperature, is concentrated in vacuo and by residue distribution in water Between (30mL) and EtOAc (50mL).Separate organic layer and through anhydrous sodium sulfate drying, filter and filtrate evaporation is obtained rough Compound.Crude product is through flash column chromatography (neutral alumina, eluant, eluent：70% ethyl acetate/hexane), obtain (4S)- N- (6- (3- hydroxy propyloxy groups) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides (0.115g, 0.226mmol, 17.25% yield), it is canescence Solid.LCMS(m/z):500.12[M+H]⁺,R_t=2.41min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.01 (s, 1H), 8.28 (d, J=7.45Hz, 1H), 8.04 (s, 1H), 7.66-7.70 (m, 2H), 7.62-7.65 (m, 1H), 7.59-7.61 (m, 1H), 7.55-7.58 (m, 1H), 7.34 (d, J= 7.89Hz, 1H), 6.42 (dd, J=7.89,0.66Hz, 1H), 5.70 (dd, J=5.81,3.18Hz, 1H), 4.21-4.15 (m, 2H), 3.65 (q, J=5.85Hz, 2H), 3.36-3.12 (m, 3H), 3.01 (dd, J=12.06,3.29Hz, 1H), 2.33 (dddd, J=14.09,9.87,5.86,4.17Hz, 1H), 2.16-2.01 (m, 2H), 1.82 (quin, J=5.92Hz, 2H).

Embodiment 313

Synthesize (4S)-N- (1- methylcyclopropyl groups) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-carboxamide hydrochlorides:

Room temperature to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diazaTriethylamine is added in the solution of (300mg, 0.983mmol) in tetrahydrofuran (20mL) (0.822mL, 5.90mmol) and triphosgene (292mg, 0.983mmol) and stirring 30min.Then 1- methyl cyclopropylamines are added (140mg, 1.965mmol) and heat 15h at 80 DEG C.(TLC systems:5%MeOH/DCM, Rf:0.4).Reactant mixture is cold But to room temperature, it is concentrated in vacuo and by residue distribution between water (25mL) and EtOAc (70mL).Separate organic layer and through anhydrous Sodium sulphate is dried, and filters and filtrate evaporation is obtained into crude compound.Thick material through flash column chromatography (100-200 silica gel, Eluant, eluent：4% ethanol/methylene), obtain colloid substance.The colloid substance is dissolved in ether (10mL) and 2.0M is added HCl (3mL) diethyl ether solution.2h is stirred at room temperature in reactant mixture and is concentrated.Gained solid is washed with ether, is expected Product (4S)-N- (1- methylcyclopropyl groups) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 5 (2H)-carboxamide hydrochlorides (170mg, 0.369mmol, 37.6% yield), it is greyish white Color solid.LCMS(m/z):403.13,[M+H]⁺, Rt=2.49min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 10.44 (s, 1H), 8.21 (d, J=7.89Hz, 1H), 8.14 (s, 1H), 8.00-7.75 (m, 2H), 7.71 (d, J=8.11Hz, 1H), 5.50 (dd, J=5.81,2.96Hz, 1H), 5.15-4.87 (m,1H),4.87-4.63(m,1H),3.56-3.29(m,4H),2.43-2.21(m,1H),2.19–1.94(m,1H),1.39 (s,3H),0.79-0.59(m,2H),0.08-0.07(m,2H).

Embodiment 314

(4S)-N- (6- ((S) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (2- picoline -4- bases) -3 is synthesized, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

0 DEG C to (4S)-N- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrazine - 2- yls) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 In the solution of stirring of (the 2H)-formamide (150mg, 0.298mmol) in methanol (3mL) be added dropwise hydrochloric acid (1mL, 32.9mmol), 5min time is lasted, 2h (TLC systems are then stirred at room temperature:5% methanol/DCM.Rf values：0.3), evaporate Solvent, is neutralized with sodium bicarbonate solution and filters gained solid, be washed with water and dry, obtain crude compound.Rough chemical combination Thing is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：2%MeOH/CH₂Cl₂), obtain desired product (4S)-N- (6- ((S) -2,3- dihydroxy propoxyl group) pyrazine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (90mg, 0.193mmol, 64.9% yield), it is light brown Solid.LCMS(m/z):464.15[M+H]⁺, Rt=1.26min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.14 (s, 1H), 8.97 (s, 1H), 8.65 (d, J=5.04Hz, 1H), 8.02 (s, 1H), 7.90-7.70 (m, 4H), 5.52 (dd, J=5.59,2.74Hz, 1H), 4.99 (d, J=4.82Hz, 1H), 4.65 (t, J=5.59Hz, 1H), 4.27-4.10 (m, 2H), 3.87-3.71 (m, 1H), 3.41 (d, J=11.40Hz, 2H), 3.25-3.03 (m, 3H), 2.97 (dd, J=12.06,3.07Hz, 1H), 2.57 (s, 3H), 2.36-2.17 (m, 1H), 2.04-1.85(m,1H)。

Embodiment 315

(4S)-N- (6- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (2- picoline -4- bases) -3 is synthesized, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

0 DEG C to (4S)-N- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine - 2- yls) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 In the solution of stirring of (the 2H)-formamide (350mg, 0.696mmol) in methanol (10mL) be added dropwise hydrochloric acid (2mL, 65.8mmol), 5min time is lasted.Then 2h is stirred at room temperature in reactant mixture.(TLC eluant, eluents：10%MeOH/ DCM:R_f- 0.3), then evaporation solvent, is neutralized with sodium bicarbonate solution, the solid of acquisition is filtered and is washed with water, and is dried, Obtain crude compound.Crude compound is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：2% methanol/DCM), Obtain desired compound (4S)-N- (6- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (210mg, 0.451mmol, 64.8% yield), it is faint yellow solid.LCMS(m/z):463.11[M+H]⁺, Rt=1.38min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 12.82 (s, 1H), 8.65 (d, J=5.26Hz, 1H), 7.92 (dd, J =5.26,1.32Hz, 1H), 7.85 (s, 1H), 7.79-7.63 (m, 4H), 6.60-6.43 (m, 1H), 5.57-5.46 (m, 1H), 4.88 (br s, 1H), 4.58 (br s, 1H), 4.18-3.99 (m, 2H), 3.83-3.67 (m, 1H), 3.38 (t, J=5.37Hz, 2H), 3.25-3.03 (m, 3H), 2.95 (dd, J=11.95,3.18Hz, 1H), 2.57 (s, 3H), 2.23-2.23 (m, 1H), 1.84-2.02(m,1H)。

Embodiment 316

(4S)-N- (4- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (2- picoline -4- bases) -3 is synthesized, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

0 DEG C to (4S)-N- (4- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyridine - 2- yls) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 In the solution of stirring of (the 2H)-formamide (200mg, 0.398mmol) in methanol (5mL) be added dropwise hydrochloric acid (1mL, 32.9mmol), 5min time is lasted.Then 2h (TLC eluant, eluents are stirred at room temperature in reactant mixture：10%MeOH/DCM: R_f- 0.2), and evaporation solvent, neutralized with sodium bicarbonate solution, the solid of acquisition is filtered and is washed with water, dried, obtain thick Produced compounds.Crude compound is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：4% methanol/DCM), it must expire Product (4S)-N- (6- ((S) -2,3- dihydroxy propoxyl group) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- two of prestige Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamide (210mg, 0.451mmol, 64.8% Yield), it is faint yellow solid.LCMS(m/z):463.18[M+H]⁺, Rt=1.13min.

¹H NMR(400MHz,DMSO-d₆):δppm 13.48(s,1H),8.58(s,1H),8.31-8.16(m,2H), 7.95 (d, J=4.82Hz, 1H), 7.83-7.65 (m, 3H), 6.74 (dd, J=5.70,2.19Hz, 1H), 5.56-5.44 (m, 1H), 5.07-4.97 (m, 1H), 4.71 (br s, 1H), 4.19-4.04 (m, 1H), 3.97 (dd, J=9.87,6.36Hz, 1H), 3.84 (br s, 1H), 3.47 (brs, 2H), 3.03-3.39 (m, 3H), 2.96 (dd, J=11.84,2.85Hz, 1H), 2.6 (s, 3H),2.33-2.12(m,1H),2.01-1.85(m,1H)。

Embodiment 317

Synthesize (4S)-N- (4- cyclopropyl pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4S)-N- (4- bromopyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (200mg, 0.485mmol), 2- cyclopropyl -4,4,5,5- tetramethyls Base -1,3,2- dioxaborolans alkane (122mg, 0.727mmol) and tripotassium phosphate (308mg, 1.455mmol) are in 1,4- bis- In oxane (4mL) and water (1mL) PdCl is added in the solution of degassing₂(dppf)(35.5mg,0.048mmol).By reaction mixing Thing stirs 16h (TLC eluant, eluents at 80 DEG C：5%MeOH/DCM:R_f-0.3).Reactant mixture is diluted with water and acetic acid second is used Ester extracts (2x10mL).The organic layer of merging salt water washing (10mL) solution, then through anhydrous sodium sulfate drying, filters and subtracts Pressure concentration, obtains crude compound.Crude compound is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：2% MeOH/ ethyl acetate), obtain desired product (4S)-N- (4- cyclopropyl pyridine -2- bases) -7- (2- picoline -4- bases) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (70mg, 0.169mmol, 34.8% yield), it is pale solid.LCMS(m/z):413.14[M+H]⁺, Rt=1.57min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.46 (s, 1H), 8.61 (d, J=5.48Hz, 1H), 8.20 (d, J= 5.92Hz, 2H), 7.95 (s, 1H), 7.71 (dd, J=5.26,1.53Hz, 1H), 7.61 (d, J=7.89Hz, 1H), 7.47 (d, J=7.89Hz, 1H), 6.71 (dd, J=5.15,1.64Hz, 1H), 5.70 (dd, J=5.81,3.18Hz, 1H), 3.42-3.09 (m, 3H), 3.07-2.91 (m, 1H), 2.67 (s, 3H) 2.42-2.23 (m, 1H), 2.09 (dt, J=14.03,6.80Hz, 1H), 1.99-1.79(m,1H),1.15-0.95(m,2H),0.93-0.72(m,2H)。

Embodiment 318

Synthesis (4S)-N- (6- (2- hydroxyl-oxethyls) pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S)-N- (6- (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethyoxyl) pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides HCl (1mL, 2.74mmol), Ran Hou are added in the solution of the stirring of (200mg, 0.351mmol) in tetrahydrofuran (15mL) 28 DEG C of stirring 1h.(TLC systems:Net ethyl acetate, Rf:0.2).Reactant mixture is concentrated in vacuo and by residue saturation NaHCO₃Solution is neutralized, and is filtered the solid obtained, is washed with water (20mLx3) and pentane (10mL × 2), obtain (4S)-N- (6- (2- hydroxyl-oxethyls) pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (98mg, 0.201mmol, 57.4% yield), it is pale solid. LCMS(m/z):487.12[M+H]⁺, Rt=2.15min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.20 (s, 1H), 9.04 (s, 1H), 8.20 (d, J=7.67Hz, 1H), 8.04 (s, 1H), 7.97 (s, 1H), 7.72-7.68 (m, 1H), 7.66-7.60 (m, 2H), 7.37 (d, J=7.89Hz, 1H), 5.71 (dd, J=5.92,3.29Hz, 1H), 4.18 (td, J=4.60,1.75Hz, 2H), 3.87-3.80 (m, 2H), 3.34- 3.14 (m, 3H), 3.03 (dd, J=12.17,3.18Hz, 1H), 2.35 (dddd, J=14.17,9.95,5.92,4.17Hz, 1H),2.14-2.00(m,2H)。

Embodiment 319

Synthesis (4S)-N- (2- (2- hydroxyl-oxethyls) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S)-N- (2- (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethyoxyl) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides HCl (1mL, 2.74mmol) is added in the solution of the stirring of (200mg, 0.351mmol) in THF (15mL), then in room temperature Stir 1h.(TLC systems:Net ethyl acetate, Rf:0.2).Reactant mixture is concentrated in vacuo and residue is filtered and uses water (20mLx3) and pentane (10mL × 2) are washed, and obtain desired product (4S)-N- (2- (2- hydroxyl-oxethyls) pyrimidine -4- Base) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (150mg, 0.305mmol, 87% yield), it is pale solid.LCMS(m/z):487.12[M+H]⁺, Rt=2.15min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.64 (s, 1H), 8.40 (d, J=6.80Hz, 1H), 8.36 (d, J= 5.70Hz, 1H), 8.08 (s, 1H), 7.82 (d, J=5.70Hz, 1H), 7.74-7.64 (m, 3H), 7.42 (d, J=8.11Hz, 1H), 5.67 (dd, J=5.92,3.07Hz, 1H), 4.40-4.37 (m, 2H), 3.94-3.90 (m, 2H), 3.34-3.15 (m, 3H), 3.02 (dd, J=12.06,3.29Hz, 1H), 2.74 (s, 1H), 2.40-2.30 (m, 1H), 2.08 (dt, J=14.14, 6.96Hz,1H)。

Embodiment 320

Synthesis (4S)-N- (6- (3- hydroxy propyloxy groups) pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Under a nitrogen in room temperature to ((4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diazaAdd in the solution of the stirring of (300mg, 0.983mmol) in tetrahydrofuran (20mL) Enter triphosgene (146mg, 0.491mmol) and stirring 30min.Into the reactant mixture add triethylamine (0.685mL, 4.91mmol) with 3- ((6- Aminopyrazine -2- bases) epoxide) propyl- 1- alcohol (216mg, 1.277mmol), then in 80 DEG C of stirrings 15h.(TLC systems:5% methanol/ethyl acetate.Rf values：0.5.).(20mL) is diluted with water in reactant mixture and acetic acid second is used Ester extracts (2x50ml).The organic layer of merging through anhydrous sodium sulfate drying and is concentrated under reduced pressure with aqueous salt solu-tion (20mL), is obtained Obtain crude compound.Through flash column chromatography, (100-200 silica gel, uses 15%MeOH/CH to crude product₂Cl₂Elution), it must expire Product (4S)-N- (6- (3- hydroxy propyloxy groups) pyrazine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros -1,4- of prestige Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (110mg, 0.212mmol, 21.56% yield), It is pale solid.LCMS(m/z):501.16[M+H]⁺, Rt=2.22min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.19 (s, 1H), 9.01 (s, 1H), 8.22 (d, J=7.67Hz, 1H), 8.03 (s, 1H), 7.93 (s, 1H), 7.73-7.58 (m, 3H), 7.37 (d, J=8.11Hz, 1H), 5.71 (dd, J=5.92, 3.29Hz, 1H), 4.27-4.14 (m, 2H), 3.72 (q, J=5.70Hz, 2H), 3.36-3.13 (m, 3H), 3.03 (dd, J= 12.06,3.29Hz, 1H), 2.41-2.29 (m, 1H), 2.10 (dt, J=14.25,6.91Hz, 1H), 1.90 (quin, J= 5.97Hz, 2H), 1.69 (t, J=5.26Hz, 1H).

Embodiment 321

Synthesize (4S)-N- (2- (3- hydroxyl -2,2- dimethyl propylenes epoxide) pyrimidine-4-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Under a nitrogen to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3-b] [1,4] diazaTriphosgene is added in the solution of (400mg, 1.310mmol) in tetrahydrofuran (20mL) (194mg, 0.655mmol) and TEA (0.913mL, 6.55mmol), is then stirred at room temperature 30min.Then, by 3- ((4- ammonia Yl pyrimidines -2- bases) epoxide) -2,2- dimethyl propylene -1- alcohol (336mg, 1.703mmol) adds to the reactant mixture, then 80 DEG C stirring 15h.(TLC systems:Ethyl acetate.Rf values：0.4).(20mL) is diluted with water in reactant mixture and ethyl acetate is used Extract (2x50ml).The organic layer of merging through anhydrous sodium sulfate drying and is concentrated under reduced pressure with aqueous salt solu-tion (20mL), is obtained Crude compound.Crude product is expected through flash column chromatography (100-200 silica gel is eluted with 0-15% methanol/DCM) Compound (4S)-N- (2- (3- hydroxyl -2,2- dimethyl propylenes epoxide) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (60mg, 0.112mmol, 8.55% yield), it is pale solid.LCMS(m/z):529.25[M+H]⁺, Rt=2.47min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.74 (s, 1H), 8.53 (br d, J=7.02Hz, 1H), 8.34 (d, J =5.70Hz, 1H), 8.06 (s, 1H), 7.78-7.69 (m, 3H), 7.65 (d, J=7.89Hz, 1H), 7.45 (d, J= 8.11Hz, 1H), 5.66 (dd, J=5.92,3.07Hz, 1H), 4.15 (s, 2H), 3.50-3.44 (m, 3H), 3.36 (s, 3H), 3.02 (dd, J=12.17,3.18Hz, 1H), 2.40-2.28 (m, 1H), 2.07 (dt, J=14.47,7.45Hz, 1H), 1.07 (s, J=16.50,9.50Hz, 6H).

Embodiment 322

Synthesize (4S) -8- chloro- N- (6- ((S) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (3- (trifluoromethyl) benzene Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-carboxamide hydrochlorides

At 0 DEG C to the chloro- N- of (4S) -8- (6- ((S) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (3- (fluoroforms Base) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (150mg, 2M HCl diethyl ether solution (2mL, 65.8mmol) 0.272mmol) is added in the suspension in ether (2mL) and gained is mixed 3h is stirred at room temperature in compound.Reactive material is stood into 5min, then solid matter sedimentation drains solvent.The solid second of acquisition Ether is ground, and is filtered and is dried in vacuo, obtains the chloro- N- of (4S) -8- (6- ((S) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine hydrochlorate (145mg, 0.243mmol, 89% yield), it is white solid.LCMS(m/z):551.05[M+H]⁺, Rt= 2.36min。

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.00 (s, 1H), 8.31 (d, J=0.88Hz, 1H), 8.09-8.18 (m, 2H), 7.97 (s, 1H), 7.90 (d, J=7.67Hz, 1H), 7.77-7.85 (m, 1H), 7.38 (d, J=0.88Hz, 1H), 5.48 (dd, J=5.81,3.18Hz, 1H), 4.30-4.39 (m, 1H), 4.20 (dd, J=10.85,6.47Hz, 1H), 3.73- 3.86 (m, 1H), 3.30-3.49 (m, 3H), 3.16-3.28 (m, 2H), 3.10 (dd, J=11.95,3.18Hz, 1H), 2.23- 2.37(m,1H),1.96-2.10(m,1H)。

Embodiment 323

(4S)-N- (2- ((S) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (2- picoline -4- bases) -3 is synthesized, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamide

0 DEG C to (4S)-N- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine - 4- yls) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 HCl (4mL, 132mmol) is added in the solution of stirring of (the 2H)-formamide (300mg, 0.596mmol) in methanol (10mL), Then 2h is stirred at room temperature.(TLC systems:5%MeOH/DCM, Rf:0.5).Reactant mixture is concentrated in vacuo, and by residue Use NaHCO₃The aqueous solution is neutralized and filters the solid of acquisition, and (2x10mL) is then washed with pentane, desired product is obtained (4S)-N- (2- ((S) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (2- picoline -4- bases) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (83mg, 0.175mmol, 29.4% yield), its For pale solid.LCMS(m/z):464.12[M+H]⁺,Rt:1.28min。

¹H NMR(400MHz,CDCl₃):δ ppm 13.67 (s, 1H), 8.68 (d, J=5.26Hz, 1H), 8.36 (d, J= 5.70Hz, 1H), 7.85 (d, J=5.70Hz, 1H), 7.79-7.74 (m, 2H), 7.65 (d, J=7.89Hz, 1H), 7.46 (d, J =7.89Hz, 1H), 5.66 (dd, J=5.59,2.96Hz, 1H), 4.44-4.31 (m, 2H), 4.14-4.02 (m, 1H), 3.77- 3.63 (m, 2H), 3.44-3.12 (m, 5H), 3.02 (dd, J=12.17,3.18Hz, 1H), 2.72 (s, 3H), 2.41-2.29 (m, 1H), 2.07 (dt, J=14.25,7.34Hz, 1H).

Embodiment 324

Synthesize (4S) -7- (6- aminopyridine -3- bases)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (20g, 63.1mmol), K₃PO₄(40.2g, 189mmol) and 5- (4,4,5,5- tetramethyl -1,3, 2- dioxaborolan -2- bases) pyridine -2- amine (16.68g, 76mmol) is in 1,4- dioxanes (400mL)/water (133mL) Pd is added in the solution of degassing₂(dba)₃(2.89g, 3.16mmol) and X-phos (3.01g, 6.31mmol).By reactant mixture Deaerate 10min again, then stirs 16h at 110 DEG C.(TLC systems:5%MeOH/ ethyl acetate, Rf:0.3).By reaction mixing Thing is cooled to room temperature, is concentrated in vacuo and is extracted residue diluted with water (500mL) (2x400mL) and with EtOAc.What is merged has Machine layer filters through anhydrous sodium sulfate drying and evaporates filtrate, obtains crude compound.Crude product is through flash column chromatography (100-200 silica gel, eluant, eluent：Net ethyl acetate), obtain desired product (4S) -7- (6- aminopyridine -3- bases)-N- (pyrroles Piperazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (14.3g, 38.2mmol, 60.5% yield), it is pale solid.LCMS(m/z):375.12[M+H]⁺, Rt=1.27min.

¹H NMR(400MHz,DMSO-d6):δ ppm 13.80 (s, 1H), 9.40 (d, J=1.31Hz, 1H), 8.72 (d, J =2.41Hz, 1H), 8.41-8.31 (m, 2H), 8.17 (dd, J=8.77,2.63Hz, 1H), 7.59 (d, J=8.11Hz, 1H), 7.49 (d, J=8.11Hz, 1H), 6.58 (d, J=8.77Hz, 1H), 6.35 (s, 2H), 5.50 (dd, J=5.92,3.07Hz, 1H), 3.24-3.01 (m, 3H), 2.93 (dd, J=11.95,3.18Hz, 1H), 2.23 (dddd, J=13.59,9.81,5.97, 3.73Hz, 1H), 1.95 (td, J=14.25,7.23Hz, 1H).

Embodiment 325

Synthesize (4S)-N- (4- (1,3- dimethyl -1H- pyrazoles -4- bases) pyridine -2- bases) -7- (2- (trifluoromethyl) pyrroles Pyridine -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

It is sub- to (4S)-N- (4- bromopyridine -2- bases) -7- (2- (trifluoromethyl) pyridin-4-yl) -3,4- dihydro -1,4- bridges Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (500mg, 0.990mmol) are in 1,4- dioxanes 1,3- dimethyl -4- (4,4,5,5- tetramethyl -1,3,2- dioxies are added in (16.0mL) and water (2.0mL) in the solution of degassing The miscellaneous amyl- 2- yls of boron heterocycle) -1H- pyrazoles (330mg, 1.484mmol), tripotassium phosphate (315mg, 1.484mmol) and PdCl₂ (dppf)-DCM(81mg,0.099mmol).Then reactant mixture is stirred into 16h at 110 DEG C.(TLC eluant, eluents：100% second Acetoacetic ester Rf:0.1；UV activation), reactant mixture is then cooled to room temperature and is diluted with water (50mL), ethyl acetate is used Extract (2 × 50mL).The organic layer of merging is filtered and concentrated through anhydrous sodium sulfate drying, obtains crude compound.Rough chemical combination Thing is through flash column chromatography (silica gel：100-200 mesh：Eluant, eluent：80% ethyl acetate/petroleum ether), then carry out preparative HPLC (preparation HPLC conditions：MP-A:10Mm ammonium hydrogen carbonate (aqueous solution) MP-B：Acetonitrile post：Kinetex c8(150*30) Mm, 5um method：50:50 flow velocitys：30ml/min；Eluant, eluent：THF+ACN+MEOH), desired product (4S)-N- (4- are obtained (1,3- dimethyl -1H- pyrazoles -4- bases) pyridine -2- bases) -7- (2- (trifluoromethyl) pyridin-4-yl) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (180mg, 0.342mmol, 34.6% yield), its For Light brown solid.LCMS(m/z):521.1[M+H]⁺,R_t=3.75min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.38 (s, 1H), 8.87 (d, J=5.04Hz, 1H), 8.48 (s, 1H), 8.33-8.25 (m, 3H), 7.70-7.65 (m, 2H), 7.52 (d, J=7.89Hz, 1H), 7.06 (dd, J=5.26,1.53Hz, 1H), 5.72 (dd, J=6.03,3.18Hz, 1H), 3.89 (s, 3H), 3.32-3.15 (m, 3H), 3.04 (dd, J=12.06, 3.29Hz, 1H), 2.52 (s, 3H), 2.40-2.31 (m, 1H), 2.11 (dt, J=14.09,7.10Hz, 1H).

Embodiment 326

Synthesis (4S)-N- (6- (2- hydroxyl-oxethyls) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S)-N- (6- (2- ((tetrahydrochysene -2H- pyrans -2- bases) epoxide) ethyoxyl) pyridine -2- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides In the solution of (250mg, 0.439mmol) in methanol (10mL) add HCl/water solution (0.6mL, 19.75mmol, 36%) and Stir 1h.(TLC eluant, eluents：100% ethyl acetate:R_f-0.2；UV activation).Reactant mixture is molten with saturated sodium bicarbonate Simultaneously solvent is evaporated under reduced pressure in basified (until pH-8-9).By residue diluted with water (10mL) and it is extracted into dichloromethane (2x20mL).The organic extract of merging filters through anhydrous sodium sulfate drying and evaporates filtrate, obtain crude compound.Slightly Material is ground and is dried under reduced pressure with 10% ethyl acetate/hexane, obtain (4S)-N- (6- (2- hydroxyl-oxethyls) pyridine -2- bases) - 7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first Acid amides (195mg, 0.401mmol, 91% yield), it is pale solid.LCMS(m/z):486.12[M+H]⁺,R_t= 2.33min。

¹H NMR(400MHz,CDCl₃):δ ppm 12.99 (s, 1H), 8.26 (d, J=7.67Hz, 1H), 8.04 (s, 1H), 7.74-7.53 (m, 5H), 7.34 (d, J=7.89Hz, 1H), 6.46 (d, J=7.89Hz, 1H), 5.70 (dd, J=5.92, 3.07Hz, 1H), 4.20-4.07 (m, 2H), 3.75 (br s, 2H), 3.35-3.12 (m, 3H), 3.02 (dd, J=12.06, 3.07Hz, 1H), 2.43-2.19 (m, 2H), 2.10 (dt, J=14.03,7.02Hz, 1H).

Embodiment 327

(4S)-N- (1- methyl -6- oxo -1,6- dihydro-pyrimidin -4- bases) -7- (3- (trifluoromethyl) phenyl) -3 is synthesized, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Under a nitrogen in room temperature to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diazaTriphosgene is added in the solution of (500mg, 1.638mmol) in THF (20mL) (340mg, 1.146mmol) and DIPEA (0.858mL, 4.91mmol) simultaneously stir 30min, then add 6- amino -3- methyl phonetic Simultaneously 24h is stirred at room temperature in reactant mixture by pyridine -4 (3H) -one (246mg, 1.965mmol).(TLC eluant, eluents：10% methanol/ DCM,Rf:0.4).By reactant mixture distribution between water (50mL) and EtOAc (100mL), organic layer is separated and through anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain crude compound.Thick material is through combiflash chromatogram purifications (silicagel column, 5% MeOH/DCM), obtain (4S)-N- (1- methyl -6- oxo -1,6- dihydro-pyrimidin -4- bases) -7- (3- (trifluoromethyl) phenyl) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (170mg, 0.369mmol, 22.53% yield), it is pale solid.LCMS(m/z):457.13[M+H]⁺, Rt=2.12min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.37 (s, 1H), 8.42 (s, 1H), 8.14 (m, J=7.89Hz, 1H), 7.92 (s, 1H), 7.70 (br d, J=7.67Hz, 1H), 7.57-7.67 (m, 2H), 7.28-7.46 (m, 1H), 7.23 (s, 1H), b5.68 (dd, J=5.92,3.29Hz, 1H), 3.49 (s, 3H), 3.09-3.34 (m, 3H), 3.00 (dd, J=12.06, 3.07Hz,1H),2.20-2.44(m,1H),1.94-2.19(m,1H)。

Embodiment 328

(4S)-N- (4- (1- methyl isophthalic acid H- pyrazoles -4- bases) pyridine -2- bases) -7- (2- picoline -4- bases) -3 is synthesized, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

It is sub- to (4S)-N- (4- bromopyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridges in room temperature Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (200mg, 0.443mmol), K₃PO₄(282mg, 1.329mmol), 1- methyl -4- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) -1H- pyrazoles (138mg, 0.665mmol) PdCl is added in 1,4- dioxanes (8mL) and water (2mL) in the solution of degassing₂(dppf)(32.4mg, 0.044mmol).Then reactant mixture is deaerated 5min again, then stirs 16h at 80 DEG C.(TLC systems:(10%MeOH/ DCM,Rf:0.3) reactant mixture, is made to be cooled to room temperature and be diluted with water.Then compound is extracted with ethyl acetate (3X20mL).The organic layer of merging is with salt water washing (2x5mL), through anhydrous sodium sulfate drying, filters and concentrates and passes through compound Flash column chromatography (100-200 mesh, eluant, eluent：Net ethyl acetate -2%DCM/MeOH) and further pass through preparation HPLC Purify (mobile phase:5mM ammonium hydrogen carbonate B:ACN methods：0/10-2/25/10/55 eluant, eluents:MeOH, THF), obtain desired production Thing (4S)-N- (4- (1- methyl isophthalic acid H- pyrazoles -4- bases) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (60mg, 0.131mmol, 29.5% yield), its For yellow solid.LCMS(m/z):453.17 [M+H]+, Rt=1.41min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.56 (s, 1H), 8.62 (d, J=5.48Hz, 1H), 8.37 (d, J= 0.66Hz, 1H), 8.31 (d, J=5.04Hz, 1H), 8.22 (s, 1H), 7.91 (d, J=0.66Hz, 1H), 7.82 (s, 1H), 7.72 (dd, J=5.37,1.43Hz, 1H), 7.63 (d, J=7.89Hz, 1H), 7.49 (d, J=7.89Hz, 1H), 7.11 (dd, J=5.15,1.64Hz, 1H), 5.71 (dd, J=6.03,3.18Hz, 1H), 3.96 (s, 3H), 3.35-3.16 (m, 3H), 3.03 (dd, J=12.06,3.29Hz, 1H), 2.75 (s, 3H), 2.41-2.31 (m, 1H), 2.17-2.05 (m, 1H).

Embodiment 329

Synthesize (4S)-7- (2,2- difluoros benzo [d] [1,3] Dioxol-4 -yl)-N- (pyridin-3-yl)-3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamide

Under a nitrogen at 0 DEG C to (4S)-7- (2,2- difluoros benzo [d] [1,3] Dioxol-4 -yl)-2,3,4, 5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(0.4g, 1.261mmol) is at tetrahydrofuran (THF) Triethylamine (1.054mL, 7.56mmol) and triphosgene (0.374g, 1.261mmol) are added in solution in (15mL) and in room Temperature stirring 1h.Pyridine -3- amine (0.237g, 2.52mmol) is added into the reactant mixture, then 16h is stirred at 90 DEG C. (TLC systems:100% ethyl acetate.Rf values：0.5).Reactant mixture is diluted with water and is extracted with ethyl acetate (2x30ml).The organic layer aqueous salt solu-tion of merging, through anhydrous sodium sulfate drying and is concentrated under reduced pressure, and obtains rough chemical combination Thing.Crude product obtains desired production through flash column chromatography (100-200 silica gel is eluted with 50% ethyl acetate/petroleum ether) Thing (4S)-7- (2,2- difluoros benzo [d] [1,3] Dioxol-4 -yl)-N- (pyridin-3-yl)-3,4- dihydros-1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (210mg, 0.474mmol, 37.6% yield), It is Light brown solid.LCMS(m/z):438.13[M+H]⁺, Rt=1.95min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.25 (s, 1H), 8.53 (d, J=2.19Hz, 1H), 8.27 (dd, J =4.71,1.42Hz, 1H), 8.05-7.87 (m, 1H), 7.80-7.66 (m, 2H), 7.57-7.47 (m, 2H), 7.43-7.30 (m, 2H), 5.49 (dd, J=5.81,2.96Hz, 1H), 3.16-3.07 (m, 3H), 2.97 (dd, J=11.95,3.18Hz, 1H), 2.25 (J=13.67,9.89,5.92,3.84Hz, 1H), 1.96 (dt, J=14.03,7.02Hz, 1H).

Embodiment 330

Synthesize (4S) -7- (6- (methylamino) pyridin-3-yl)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamide

To the chloro- N- of (4S) -7- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-formamides (4.2g, 13.26mmol), N- methyl -5- (4,4,5,5- tetramethyl -1,3,2- dioxa boron heterocycles Amyl- 2- yls) pyridine -2- amine (3.41g, 14.59mmol) and Na₂CO₃(4.22g, 39.8mmol) is at 1,4- dioxanes (18mL): Pd (PPh are added in water (4.50mL) in the solution of degassing₃)₄(0.766g,0.663mmol).Reactant mixture is stirred at 110 DEG C Mix 16h.(TLC systems:5% methanol/ethyl acetate, Rf:0.3).So that reactant mixture is cooled to room temperature, then it is evaporated under reduced pressure 1,4- dioxane solvents, by the residue diluted with water (50mL) of acquisition and are extracted with ethyl acetate (2x100ml).What is merged has Machine layer water, salt water washing, it is dried over sodium sulfate and solvent is evaporated under reduced pressure, obtain crude product.Crude product is pure through flash column chromatography Change (silica gel 100-200 mesh：Eluant, eluent：Net ethyl acetate), obtain desired product (4S) -7- (6- (methylamino) pyridines -3- Base)-N- (pyrazine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine (2.50g, 6.40mmol, 48.3% yield), it is white solid.LCMS(m/z):389.16[M+H]⁺, Rt= 1.31min。

¹H NMR(400MHz,CDCl₃):δ ppm 13.90 (s, 1H), 9.54 (d, J=1.53Hz, 1H), 8.75 (d, J= 2.63Hz, 1H), 8.36-8.29 (m, 3H), 8.28 (s, 1H), 7.55 (d, J=7.89Hz, 1H), 6.53 (d, J=8.77Hz, 1H), 5.69 (dd, J=6.03,3.18Hz, 1H), 4.78 (d, J=4.82Hz, 1H), 3.33-3.13 (m, 3H), 3.07-2.97 (m,4H),2.35-2.26(m,1H),2.13-2.03(m,1H)。

Embodiment 331

(4S)-N- (6- ((S) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) -7- (2- picoline -4- bases) -3 is synthesized, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

0 DEG C to (4S)-N- (6- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) pyrimidine - 4- yls) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 Hydrochloric acid (1mL, 32.9mmol) is added dropwise in the solution of stirring of (the 2H)-formamide (50mg, 0.099mmol) in methanol (3mL), Last 5min time.Then, reactant mixture is stirred into 30min at 28 DEG C.(TLC eluant, eluents：10%MeOH/DCM:R_f- 0.2；UV activation).Reactant mixture is neutralized and filtered with sodium bicarbonate solution the solid obtained, is washed with water, Ran Houyong The grinding of 50% ether/pentane, obtain desired compound (4S)-N- (6- ((S) -2,3- dihydroxy propoxyl group) pyrimidine-4-yl) - 7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first Acid amides (80mg, 0.165mmol, 167% yield), it is yellow solid.LCMS(m/z):464.15[M+H]⁺,Rt: 1.26min。

¹H NMR(400MHz,DMSO-d₆):δppm 13.72(s,1H),8.62-8.54(m,2H),8.15(s,1H), 7.93 (s, 1H), 7.82 (d, J=7.45Hz, 1H), 7.74 (d, J=7.23Hz, 1H), 7.47 (s, 1H), 5.49 (s, 1H), 4.98 (d, J=4.38Hz, 1H), 4.67 (s, 1H), 4.38 (d, J=8.55Hz, 1H), 4.26-4.18 (m, 1H), 3.82 (s, 1H),3.45(s,2H),3.29(s,3H),3.14-2.91(m,1H),2.63(s,3H),2.24(s,1H),1.96(s,1H)。

Embodiment 332

Synthesis (4S)-N- (4- (2- Jia Ji oxazole -5- bases) pyridine -2- bases) -7- (2- (trifluoromethyl) pyridin-4-yl) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (4S)-N- (4- bromopyridine -2- bases) -7- (2- (trifluoromethyl) pyridin-4-yl) -3,4- dihydro -1, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (500mg, 0.990mmol) are in 1,4- bis- Evil Alkane (20mL):In water (5mL) K is added in the solution of degassing₃PO₄(630mg, 2.97mmol), 2- methyl -5- (4,4,5,5- tetramethyls Base -1,3,2- dioxaborolan -2- base) oxazoles (310mg, 1.484mmol) and PdCl₂(dppf)-CH₂Cl₂Adduct (81mg, 0.099mmol)), reactant mixture is then stirred into 16h at 85 DEG C.(TLC systems:Rf-0.2,EtOAc).Will reaction Mixture is cooled to room temperature and is diluted with water (120mL), and (2x250mL) is extracted with ethyl acetate, with salt water washing (100mL). The organic layer of merging is through anhydrous sodium sulfate drying and is concentrated under reduced pressure, and obtains crude compound.Crude product is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：95% ethyl acetate/hexane) and it is purified into (condition again by preparation HPLC：Stream Dynamic phase A:10.0mM ammonium hydrogen carbonate Mobile phase B:Acetonitrile；Constant gradient：50:50(A:B) post：Xbridge C18(250*30)mm, 10um eluant, eluents：The excessive THF+MeOH flow velocitys of acetonitrile+volume：30ml/min), desired product (4S)-N- (4- (2- are obtained Jia Ji oxazole -5- bases) pyridine -2- bases) -7- (2- (trifluoromethyl) pyridin-4-yl) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides (240mg, 0.473mmol, 47.8% yield), it is solid for canescence Body.LCMS(m/z):508.1[M+H]⁺, Rt=2.37min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.48 (s, 1H), 8.87 (d, J=5.26Hz, 1H), 8.46 (d, J= 12.72Hz, 2H), 8.36 (d, J=5.26Hz, 1H), 8.23 (dd, J=5.04,1.32Hz, 1H), 7.68 (d, J=7.89Hz, 1H), 7.53 (d, J=8.11Hz, 1H), 7.46 (s, 1H), 7.21 (dd, J=5.15,1.43Hz, 1H), 5.73 (dd, J= 5.81,3.18Hz, 1H), 3.35-3.14- (m, 3H), 3.06 (dd, J=12.06,3.29Hz, 1H), 2.56 (s, 3H), 2.47- 2.28 (m, 1H), 2.12 (dt, J=14.20,7.04Hz, 1H).

Embodiment 333

Synthesize (4S)-N- (4- (oxazole -5- bases) pyridine -2- bases) -7- (2- (trifluoromethyl) pyridin-4-yl) -3,4- two Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (4S)-N- (4- bromopyridine -2- bases) -7- (2- (trifluoromethyl) pyridin-4-yl) -3,4- dihydro -1, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (500mg, 0.990mmol) are in 1,4- bis- Evil Alkane (20mL):K is added in the solution of degassing in water (5mL)₃PO₄(630mg, 2.97mmol), 5- (4,4,5,5- tetramethyl -1, 3,2- dioxaborolan -2- base) oxazoles (289mg, 1.484mmol) and PdCl₂(dppf)-CH₂Cl₂Adduct (81mg, 0.099mmol), reactant mixture is then stirred into 16h at 85 DEG C.(TLC systems:Rf-0.2,EtOAc).Make reactant mixture It is cooled to RT and is diluted with water (120mL), is extracted with ethyl acetate (2x250mL).The organic layer of merging is dry through anhydrous sodium sulfate It is dry and be concentrated under reduced pressure, obtain crude compound.Crude product is through flash column chromatography (silica gel：100-200 mesh, eluant, eluent：90% Ethyl acetate/hexane) and it is purified into (condition again by preparation HPLC：MP-A:10mm ammonium hydrogen carbonate MP-B：Acetonitrile post： Xterra (150*19mm, 10u) sun methods：T/%B 0/25,1/25,6/65,13/65,13.10/100,16/100, 16.10/25,19/25 flow velocity：17ml/min eluant, eluents：ACN+THF+MEOH), desired product (4S)-N- (4- (Evil are obtained Azoles -5- bases) pyridine -2- bases) -7- (2- (trifluoromethyl) pyridin-4-yl) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza- 5 (2H)-formamides (210mg, 0.423mmol, 42.7% yield), it is pale solid.LCMS (m/z):494.1[M+H]⁺, Rt=8.46min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.53 (s, 1H), 8.89 (d, J=5.04Hz, 1H), 8.54-8.47 (m, 2H), 8.41 (d, J=5.26Hz, 1H), 8.22 (dd, J=5.15,1.43Hz, 1H), 8.00 (s, 1H), 7.70 (d, J= 8.11Hz, 1H), 7.62 (s, 1H), 7.54 (d, J=7.89Hz, 1H), 7.32-7.27 (m, 1H), 5.74 (dd, J=5.92, 3.07Hz, 1H), 3.37-3.16 (m, 3H), 3.07 (dd, J=12.06,3.29Hz, 1H), 2.46-2.32 (m, 1H), 2.13 (dt, J=14.09,6.88Hz, 1H).

Embodiment 334

Synthesize (4S)-N- (4- (2- Jia Ji oxazole -4- bases) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- two Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C under a nitrogen to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diazaThree light are added in the solution of the stirring of (8g, 31.7mmol) in tetrahydrofuran (600mL) Gas (5.65g, 19.02mmol) and DIPEA (28mL, 159mmol), are then stirred at room temperature 30min.Then, 4- (2- first is added Ji oxazole -5- bases) pyridine -2- amine (8.33g, 47.6mmol), then stirs 16h at 80 DEG C.(TLC:10%MeOH/ acetic acid second Ester, Rf:0.3).So that reactant mixture is cooled to room temperature, pours into cold water (150mL), be extracted with ethyl acetate (2x300mL).The organic layer of merging is through anhydrous sodium sulfate drying and is concentrated under reduced pressure, and obtains crude compound.Crude compound is passed through Flash column chromatography (neutral alumina, 65% ethyl acetate/petroleum ether), obtains desired product (6g).Absorbed (800mL) and addition Silicycle palladiums scavenger (3g), then stir 4h at 50 DEG C in ethyl acetate.By mixture through diatom Soil pad is filtered and washed (50ml) with hot ethyl acetate, and the filtrate decompression of acquisition is concentrated, and obtains (4S)-N- (4- without Pd (2- Jia Ji oxazole -4- bases) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (5.5g, 12.12mmol, 38.2% yield), it is yellow solid. LCMS(m/z):454.22[M+H]⁺,R_t=1.53min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.65 (s, 1H), 8.62 (d, J=5.26Hz, 1H), 8.47-8.52 (m, 1H), 8.38 (dd, J=5.26,0.66Hz, 1H), 8.17-8.23 (m, 1H), 7.72 (dd, J=5.15,1.64Hz, 1H), 7.63 (d, J=7.89Hz, 1H), 7.43-7.52 (m, 2H), 7.20 (dd, J=5.26,1.53Hz, 1H), 5.72 (dd, J= 5.92,3.07Hz, 1H), 3.15-3.37 (m, 3H), 3.03 (dd, J=12.06,3.29Hz, 1H), 2.75 (s, 3H), 2.56 (s, 3H), 2.36 (dddd, J=14.00,9.89,5.92,3.95Hz, 1H), 2.05-2.16 (m, 1H).

Embodiment 335

Synthesize (4S)-N- (4- (2- Jia Ji oxazole -5- bases) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- two Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-carboxamide hydrochlorides

At 0 DEG C to (4S)-N- (4- (2- Jia Ji oxazole -5- bases) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (100mg, 0.221mmol) are in second 2M HCl diethyl ether solution (2.5mL, 0.221mmol), last 2min one section are added dropwise in the solution of stirring in ether (10mL) Time.Then reactant mixture is stirred into 2h at 30 DEG C.(TLC:10%MeOH/DCM, Rf:0.2), and solvent is evaporated under reduced pressure, will The solid chemical compound of acquisition washs (2x20mL) with pentane, obtains desired compound (4S)-N- (4- (2- Jia Ji oxazoles -5- Base) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneous- 5 (2H)-carboxamide hydrochlorides (84mg, 0.171mmol, 78% yield), it is faint yellow solid.LCMS(m/z): 454.22[M+H]⁺,R_t=1.52min.

¹H NMR(400MHz,CD₃OD):δ ppm 8.93 (d, J=6.14Hz, 1H), 8.69 (s, 1H), 8.59 (d, J= 5.92Hz, 1H), 8.45 (d, J=6.14Hz, 1H), 8.32 (d, J=8.11Hz, 1H), 8.14 (d, J=8.11Hz, 1H), 8.04 (s, 1H), 7.96 (s, 1H), 7.78 (d, J=6.14Hz, 1H), 5.88 (d, J=5.70Hz, 1H), 4.06-3.98 (m, 4H), 2.95 (s, 3H), 2.84-2.71 (m, 2H), 2.67 (s, 3H), 2.59 (dt, J=14.31,6.99Hz, 1H).

Embodiment 336

Synthesize (4R)-N- (4- (2- Jia Ji oxazole -5- bases) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- two Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4R)-N- (4- bromopyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (450mg, 0.997mmol), 2- methyl -5- (4,4,5,5- tetramethyls Base -1,3,2- dioxaborolan -2- base) oxazoles (313mg, 1.496mmol) and K₃PO₄(635mg, 2.99mmol) 1, 4- dioxanes (16mL):In water (4mL) PdCl is added in the solution of degassing₂(dppf)(73.0mg,0.100mmol).Will reaction Mixture stirs 16h at 90 DEG C.(TLC:10%MeOH/EtOAc Rf:0.7.1,4- dioxane solvents are evaporated under reduced pressure, will obtain Residue diluted with water (50mL) and be extracted with ethyl acetate (2x100mL).The organic layer water of merging, salt water washing, warp Sodium sulphate is dried and solvent is evaporated under reduced pressure, and obtains crude product.Crude product is through flash column chromatography ((silica gel 100-200 mesh：Wash De- agent：2%MeOH/DCM), desired product (4R)-N- (4- (2- Jia Ji oxazole -5- bases) pyridine -2- bases) -7- (2- first is obtained Yl pyridines -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (250mg, 0.550mmol, 55.1% yield), it is white solid.LCMS(m/z):454.22[M+H]⁺,R_t=1.52min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.66 (s, 1H), 8.63 (d, J=5.26Hz, 1H), 8.49 (s, 1H), 8.38 (d, J=5.26Hz, 1H), 8.19 (d, J=1.32Hz, 1H), 7.72 (dd, J=5.26,1.75Hz, 1H), 7.63 (d, J =7.89Hz, 1H), 7.52-7.45 (m, 2H), 7.21 (dd, J=5.26,1.53Hz, 1H), 5.72 (dd, J=5.92, 3.29Hz, 1H), 3.36-3.14 (m, 3H), 3.04 (dd, J=12.06,3.29Hz, 1H), 2.75 (s, 3H), 2.56 (s, 3H), 2.42-2.30 (m, 1H), 2.11 (dt, J=14.09,7.10Hz, 1H).

Embodiment 337

Synthesis (4S)-N- (4- (2- methylthiazol -5- bases) pyridine -2- bases) -7- (2- (trifluoromethyl) pyridin-4-yl) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

It is sub- to (4S)-N- (4- bromopyridine -2- bases) -7- (2- (trifluoromethyl) pyridin-4-yl) -3,4- dihydro -1,4- bridges Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (500mg, 0.990mmol) are in 1,4- dioxanes 2- methyl -5- (4,4,5,5- tetramethyl -1,3,2- dioxa boron is added in (20.0mL) and water (5.0mL) in the solution of degassing The amyl- 2- yls of heterocycle) thiazole (334mg, 1.484mmol), Na₂CO₃(315mg, 2.97mmol) and Pd (TPP)₄(57.2mg, 0.049mmol) and by reactant mixture at 110 DEG C stir 16h.(TLC systems:100% ethyl acetate, Rf values：0.3).Will be anti- Answer mixture to be cooled to room temperature, (30mL) is diluted with water and is extracted with ethyl acetate (2 × 20mL).The organic layer of merging is through anhydrous Sodium sulphate is dried, filtered and concentrated.Crude compound is through flash column chromatography (silica gel 100-200 mesh, with 40% acetic acid second The elution of ester/petroleum ether), then purify (post with preparation HPLC：Xbridge C18(50x19mm)5μ；Mobile phase-A:10M carbon Sour hydrogen ammonium；Mobile phase-B：Acetonitrile, method (T/%B):0/20,/10/55；Flow velocity：17ml/min；Eluant, eluent：Acetonitrile+MeOH+ THF+TFA), obtain (4S)-N- (4- (2- methylthiazol -5- bases) pyridine -2- bases) -7- (2- (trifluoromethyl) pyridin-4-yl) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (195mg, 0.372mmol, 37.6% yield), it is pale solid.LCMS(m/z):524.13[M+H]⁺, Rt=2.54min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.47 (s, 1H), 8.87 (d, J=5.26Hz, 1H), 8.48 (s, 1H), 8.41 (d, J=0.88Hz, 1H), 8.33 (d, J=5.26Hz, 1H), 8.22 (dd, J=5.15,1.64Hz, 1H), 8.05 (s, 1H), 7.68 (d, J=7.89Hz, 1H), 7.53 (d, J=7.89Hz, 1H), 7.14 (dd, J=5.26,1.53Hz, 1H), 5.73 (dd, J=5.92,3.07Hz, 1H), 3.36-3.15 (m, 3H), 3.05 (dd, J=12.17,3.18Hz, 1H), 2.76 (s, 3H), 2.37 (qd, J=9.90,4.28Hz, 1H), 2.12 (dt, J=14.14,7.18Hz, 1H).

Embodiment 338

Synthesize (4S)-N- (4- (1H-1,2,3- triazole -5- bases) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaAdded in the solution of the stirring of (300mg, 1.189mmol) in tetrahydrofuran (10mL, seal pipe) TEA (0.829mL, 5.94mmol) and triphosgene (353mg, 1.189mmol) and stirring 45min.Then in room temperature by 4- (1H- 1,2,3-triazoles -5- bases) pyridine -2- amine (287mg, 1.783mmol) adds to reactant mixture, then stirs 16h at 80 DEG C. (TLC systems:5%MeOH DCM.R_fValue：0.3,UV).Reactant mixture is cooled to room temperature, (15mL) is diluted with water, second is used Acetoacetic ester extracts (5X20mL).The organic layer of merging washs (10mL) with saturated brine solution, through anhydrous sodium sulfate drying, filtering And concentrate, obtain crude compound.Crude compound purifies (condition through preparation HPLC：MP-A:10Mm ammonium hydrogen carbonate is (water-soluble Liquid), MPB：Acetonitrile, post：Atlantis T3 (250*19mm*5u), method：0/35,11/35,11.1/100,15/100, 15.1/35,19/35, flow velocity：16ml/min, eluant, eluent：ACN+THF+MeOH+DMSO), desired product (4S)-N- is obtained (4- (1H-1,2,3- triazole -5- bases) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (60mg, 0.133mmol, 11.15% yield), it is canescence Solid.LCMS(m/z):440.21[M+H]⁺,R_t=1.42min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.57 (s, 1H), 8.66 (s, 1H), 8.60 (d, J=5.26Hz, 1H), 8.49-8.39 (m, 2H), 8.25 (s, 1H), 7.96 (dd, J=5.26,1.53Hz, 1H), 7.85-7.79 (m, 2H), 7.72 (d, J=7.89Hz, 1H), 7.56 (dd, J=5.15,1.43Hz, 1H), 5.54 (dd, J=5.70,3.07Hz, 1H), 3.17-3.06 (m, 3H), 2.98 (dd, J=12.06,3.29Hz, 1H), 2.64 (m, 3H), 2.36-2.20 (m, 1H), 1.98 (dt, J=13.70,6.96Hz, 1H).

Embodiment 339

(4S)-N- (5- (1- methyl isophthalic acid H- pyrazoles -4- bases) pyridin-3-yl) -7- (2- picoline -4- bases) -3 is synthesized, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

It is sub- to (4S)-N- (5- bromopyridine -3- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridges in room temperature Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (400mg, 0.886mmol), Na₂CO₃(282mg, 2.66mmol) and 1- methyl -4- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) -1H- pyrazoles (277mg, Pd (Ph 1.329mmol) are added in the solution of degassing in 1,4- dioxanes (28mL) and water (7.00mL)₃P)₄(51.2mg, 0.044mmol) and by reactant mixture at 75 DEG C stir 16h.(TLC system 10%MeOH/ ethyl acetate, Rf:0.2).Will be anti- Mixture is answered to be diluted with water and be extracted with ethyl acetate (2x100mL).The organic layer water and salt water washing of merging, through Na₂SO₄ It is dried, filtered and concentrated, obtains crude compound.Crude product is through flash column chromatography (neutral alumina, eluant, eluent：2% MeOH/EtOAc), desired product (4S)-N- (5- (1- methyl isophthalic acid H- pyrazoles -4- bases) pyridin-3-yl) -7- (2- methyl is obtained Pyridin-4-yl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (120mg, 0.262mmol, 29.6% yield), it is white solid.LCMS(m/z):453.29[M+H]⁺,R_t=1.34min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.13 (s, 1H), 8.68 (d, J=5.04Hz, 1H), 8.46 (d, J= 1.75Hz, 1H), 8.38 (q, J=2.41Hz, 2H), 7.81 (d, J=0.66Hz, 1H), 7.67 (d, J=7.02Hz, 1H), 7.60 (s, 1H), 7.54-7.48 (m, 2H), 7.38 (d, J=7.89Hz, 1H), 5.70 (dd, J=5.92,3.07Hz, 1H), 3.96 (s, 3H), 3.35-3.16 (m, 3H), 3.04 (dd, J=12.06,3.29Hz, 1H), 2.67 (s, 3H), 2.28-2.44 (m, 1H), 2.11 (dt, J=14.09,6.88Hz, 1H).

Embodiment 340

Synthesize (4S) -7- (2- picoline -4- bases)-N- (4- (2- oxooxazolidine -3- bases) pyridine -2- bases) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaAdded in the solution of the stirring of (350mg, 1.387mmol) in tetrahydrofuran (10mL, seal pipe) TEA (0.967mL, 6.94mmol), triphosgene (412mg, 1.387mmol) and stirring 45min.Then in room temperature by 3- (2- ammonia Yl pyridines -4- base) oxazolidine -2- ketone (373mg, 2.081mmol) adds to reactant mixture, then stirs 16h at 80 DEG C.(TLC System:5%MeOH DCM.R_fValue：0.3,UV).Reactant mixture is cooled to room temperature and is diluted with water (15mL), use acetic acid second Ester extracts (5X20mL).The organic layer of merging washs (10mL) with saturated brine solution, through anhydrous sodium sulfate drying, filters and subtracts Pressure concentration filtrate, obtains crude compound.Crude compound purifies (condition through preparation HPLC：MP A:10mM ammonium hydrogen carbonate (aqueous solution), MP B：Acetonitrile, post：Kromosil C18 (250*21.2) mm 10u, method：T/%B=0/40,12/40, 12.5/100,16/100,16.5/40, flow velocity：20ml/min, eluant, eluent：Acetonitrile+THF+ water), obtain desired product (4S)- 7- (2- picoline -4- bases)-N- (4- (2- oxooxazolidine -3- bases) pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (54mg, 0.117mmol, 8.47% yield), it is canescence Solid.LCMS(m/z):458.26[M+H]⁺,R_t=1.48min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.65 (s, 1H), 8.62 (d, J=5.26Hz, 1H), 8.30 (d, J =5.70Hz, 1H), 8.20 (s, 1H), 8.06 (d, J=1.75Hz, 1H), 7.80 (dd, J=5.81,2.08Hz, 1H), 7.71 (d, J=4.17Hz, 1H), 7.63 (d, J=7.89Hz, 1H), 7.49 (d, J=7.89Hz, 1H), 5.68 (dd, J=5.70, 3.29Hz, 1H), 4.53 (t, J=8.00Hz, 2H), 4.15 (t, J=8.00Hz, 2H), 3.40-3.13 (m, 3H), 3.02 (dd, J=11.95,3.18Hz, 1H), 2.74 (s, 3H), 2.44-2.21 (m, 1H), 2.09 (dt, J=13.98,6.71Hz, 1H).

Embodiment 341

Synthesize (4S)-N- (5- (1,3- dimethyl -1H- pyrazoles -4- bases) pyridin-3-yl) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

It is sub- to (4S)-N- (5- bromopyridine -3- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridges in room temperature Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (400mg, 0.886mmol), Na₂CO₃(282mg, 2.66mmol), 1,3- dimethyl -4- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) -1H- pyrazoles (295mg, 1.329mmol) adds Pd (Ph in 1,4- dioxanes (20mL) and water (5mL) in the solution of degassing₃P)₄ (51.2mg, 0.044mmol) and the 5min that deaerates again.Then reactant mixture is stirred into 15h at 80 DEG C.(TLC systems:10% Ethanol/methylene, Rf:0.2).Reactant mixture is cooled to room temperature, dioxane is then evaporated from reactant mixture molten Agent, obtains residue, is diluted with water and is extracted with ethyl acetate (3X50mL).The organic layer of merging salt water washing (2x20mL), through anhydrous sodium sulfate drying, filters and concentrates, and obtains crude compound.Thick material is through flash column chromatography (silicon Glue:100-200 mesh, eluant, eluent：90% ethyl acetate/petroleum ether), obtain desired product (4S)-N- (5- (1,3- dimethyl- 1H- pyrazoles -4- bases) pyridin-3-yl) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza- 5 (2H)-formamides (195mg, 0.417mmol, 47.1% yield), it is pale solid.LCMS (m/z):467.1[M+H]⁺,R_t=3.27.

¹H NMR(400MHz,CDCl₃):δ ppm 13.12 (s, 1H), 8.66 (d, J=5.04Hz, 1H), 8.42 (d, J= 2.41Hz, 1H), 8.38 (d, J=1.97Hz, 1H), 8.31 (t, J=2.08Hz, 1H), 7.65 (d, J=7.89Hz, 1H), 7.60 (s, 1H), 7.53-7.49 (m, 2H), 7.38 (d, J=7.89Hz, 1H), 5.70 (dd, J=5.81,3.18Hz, 1H), 3.89 (s, 3H), 3.34-3.15 (m, 3H), 3.04 (dd, J=12.17,3.18Hz, 1H), 2.66 (s, 3H), 2.42 (s, 3H), 2.40-2.31 (m, 1H), 2.11 (dt, J=14.31,7.43Hz, 1H).

Embodiment 342

Synthesize (4S)-N- (4- (1H-1,2,3- triazol-1-yls) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaAdded in the solution of the stirring of (450mg, 1.783mmol) in tetrahydrofuran (10mL, seal pipe) TEA (1.243mL, 8.92mmol) and triphosgene (529mg, 1.783mmol) and stirring 45min.Then at 28 DEG C by 4- (1H- 1,2,3-triazoles -1- bases) pyridine -2- amine (345mg, 2.140mmol) adds to reactant mixture, then stirs 4h at 80 DEG C. (TLC systems:5%MeOH DCM.R_fValue：0.3,UV).Reactant mixture is cooled to room temperature and is diluted with water (15mL), use second Acetoacetic ester extracts (5X20mL).The organic layer of merging washs (10mL) with saturated brine solution, through anhydrous sodium sulfate drying, filtering And concentrate, obtain crude compound.Crude compound purifies (condition through preparation HPLC：MP-A:10mM ammonium hydrogen carbonate is (water-soluble Liquid) MP-B：Acetonitrile, post：The μ of kromasil (2150*21.1mm) 5, method：0/10,1/10,10/55, flow velocity：17ml/min, is washed De- agent：ACN+MeOH), desired product (4S)-N- (4- (1H-1,2,3- triazol-1-yls) pyridine -2- bases) -7- (2- first is obtained Yl pyridines -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (175mg, 0.398mmol, 22.32% yield), it is pale solid.LCMS(m/z):440.22[M+H]⁺,R_t= 1.60min。

¹H NMR(400MHz,DMSO-d₆):δ ppm 13.83 (s, 1H), 9.03 (d, J=1.10Hz, 1H), 8.81 (d, J =1.53Hz, 1H), 8.61 (t, J=5.81Hz, 2H), 8.21 (s, 1H), 8.07 (d, J=1.32Hz, 1H), 7.95 (dd, J= 5.26,1.53Hz, 1H), 7.87-7.79 (m, 1H), 7.79-7.62 (m, 1H), 5.54 (dd, J=5.92,3.07Hz, 1H), 3.28-3.19 (m, 1H), 3.18-3.06 (m, 3H), 2.99 (dd, J=11.95,3.18Hz, 1H), 2.62 (s, 3H), 2.48- 2.20(m,1H),2.09-1.85(m,1H)。

Embodiment 343

Synthesize (4S) -7- (2- picoline -4- bases)-N- (5- (2- methylthiazol -5- bases) pyridin-3-yl) -3,4- two Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

It is sub- to (4S)-N- (5- bromopyridine -3- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridges in room temperature Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (500mg, 1.108mmol), K₃PO₄(705mg, 3.32mmol), 2- methyl -5- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) thiazole (374mg, 1.662mmol) PdCl is added in 1,4- dioxanes (16mL) and water (4mL) in the solution of degassing₂(dppf)-CH₂Cl₂Adduction Thing (136mg, 0.166mmol), then deaerates reactant mixture 5min again, then stirs 15h at 80 DEG C.(TLC systems: 10%MeOH/DCM, Rf:0.2).Reactant mixture is cooled to room temperature and evaporation reactant mixture, crude product is obtained.Roughization Compound obtains desired product (4S) -7- (2- through flash column chromatography (neutral alumina, 80% ethyl acetate/petroleum ether) Picoline -4- bases)-N- (5- (2- methylthiazol -5- bases) pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (190mg, 0.393mmol, 35.4% yield), it is brown solid. LCMS(m/z):470.24[M+H]⁺,R_t=1.59min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.21 (s, 1H), 8.69 (d, J=5.04Hz, 1H), 8.52-8.43 (m, 3H), 7.88 (s, 1H), 7.66 (d, J=7.89Hz, 1H), 7.58 (s, 1H), 7.51 (dd, J=5.15,1.21Hz, 1H), 7.39 (d, J=7.89Hz, 1H), 5.70 (dd, J=5.81,3.18Hz, 1H), 3.15-3.36 (m, 3H), 3.04 (dd, J= 12.06,3.29Hz, 1H), 2.76 (s, 3H), 2.67 (s, 3H), 2.36 (qd, J=10.01,4.17Hz, 1H), 2.11 (dt, J =13.98,6.93Hz, 1H).

Embodiment 344

Synthesize (4S)-N- (4- (1H-1,2,4- triazole -5- bases) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaAdded in the solution of the stirring of (450mg, 1.783mmol) in tetrahydrofuran (10mL, seal pipe) TEA (1.243mL, 8.92mmol) and triphosgene (529mg, 1.783mmol), are then stirred at room temperature 45min.Then in room temperature 4- (1H-1,2,4- triazole -5- bases) pyridine -2- amine (345mg, 2.140mmol) is added into reactant mixture, then stirred at 80 DEG C Mix 16h.(TLC systems:5%MeOH DCM.R_fValue：0.3,UV).Reactant mixture is set to be cooled to room temperature and be diluted with water (15mL), is extracted with ethyl acetate (5X20mL).The organic layer of merging washs (10mL) with saturated brine solution and through anhydrous Na₂SO₄Dry, filter and concentrate filtrate decompression, obtain crude compound.Crude compound is purified through preparation HPLC and (prepared Type HPLC methods：MP-A:10mM ammonium hydrogen carbonate (aqueous solution) MP-B：Acetonitrile, post：Kinetex C8 (150*30) mm, 5u, side Method：0/10,1/10,8/40,12/40,12.1/100,15/100,15.1/10, flow velocity：30ml/min, eluant, eluent：Acetonitrile+THF + methanol+DMSO+DMSO+ formic acid), obtain desired product (4S)-N- (4- (1H-1,2,4- triazole -5- bases) pyridine -2- bases) - 7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first Acid amides (102mg, 0.221mmol, 12.38% yield), it is pale solid.LCMS(m/z):440.2[M+H]⁺,R_t= 3.71min.

¹H NMR(400MHz,DMSO-d₆):δ ppm 14.52 (s, 1H), 13.63 (s, 1H), 8.84 (d, J=0.88Hz, 1H), 8.70-8.55 (m, 1H), 8.54-8.38 (m, 1H), 8.37-8.35 (d, 1H) 8.25 (s, 1H), 7.95 (dd, J= 5.37,1.43Hz, 1H), 7.86-7.77 (m, 1H), 7.77-7.64 (m, 2H), 5.54 (dd, J=5.92,3.07Hz, 1H), 3.18-3.05 (m, 3H), 2.98 (dd, J=11.95,3.18Hz, 1H), 2.78-2.52 (m, 3H), 2.48-2.36 (m, 1H), 2.07(s,1H)。

Embodiment 345

Synthesize (4S)-N- (5- (2- Jia Ji oxazole -5- bases) pyridin-3-yl) -7- (2- picoline -4- bases) -3,4- two Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

It is sub- to (4S)-N- (5- bromopyridine -3- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridges in room temperature Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (350mg, 0.776mmol), Na₂CO₃(247mg, 2.327mmol) and 2- methyl -5- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- base) oxazoles (243mg, Pd (Ph 1.163mmol) are added in the solution of the degassing in 1,4- dioxanes (20mL) and water (4mL)₃P)₄(44.8mg, Deaerate 0.039mmol) and again 5min.Then gained reactant mixture is stirred to 80 DEG C, keeps 15h.(TLC systems:10% MeOH/DCM,Rf:0.3) and cause reactant mixture to be cooled to room temperature, evaporation of organic solvent and will be obtained from reactant mixture Residue diluted with water, be extracted with ethyl acetate (3X20mL).The organic layer of merging is washed with saturated brine solution (2x5mL), through anhydrous sodium sulfate drying, filters and concentrates, and obtains thick material.Crude compound purifies (bar through preparation HPLC Part：MP-A:10mM ammonium hydrogen carbonate (aqueous solution) MP-B：Acetonitrile post：Sunfire C8 (150*19) mm, 10u method：-0/10,1/ 10,10/45 flow velocity：18ml/min；Eluant, eluent：Methanol+CAN+THF), obtain desired product (4S)-N- (5- (2- Jia Ji Evil Azoles -5- bases) pyridin-3-yl) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (68mg, 0.149mmol, 19.17% yield), it is pale solid.LCMS(m/z): 454.1[M+H]⁺, Rt=3.39min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.24 (s, 1H), 8.69 (d, J=5.26Hz, 1H), 8.58 (d, J= 1.75Hz, 1H), 8.52 (d, J=2.19Hz, 1H), 8.48-8.44 (m, 1H), 7.66 (d, J=7.89Hz, 1H), 7.59 (s, 1H), 7.52 (d, J=5.26Hz, 1H), 7.39 (d, J=7.89Hz, 1H), 7.33 (s, 1H), 5.71 (dd, J=5.81, 3.18Hz, 1H), 3.37-3.15 (m, 3H), 3.05 (dd, J=12.17,3.40Hz, 1H), 2.68 (s, 3H), 2.56 (s, 3H), 2.37 (qd, J=9.79,4.38Hz, 1H), 2.11 (dt, J=14.36,7.07Hz, 1H).

Embodiment 346

(4S)-N- (4- (1- methyl isophthalic acid H- pyrazoles -4- bases) pyrimidine -2-base) -7- (2- picoline -4- bases) -3 is synthesized, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In a nitrogen atmosphere room temperature in seal pipe to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysenes - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(350mg, 1.387mmol) stirring in tetrahydrofuran (25mL) Triphosgene (247mg, 0.832mmol), DIPEA (1.211mL, 6.94mmol) are added in the solution mixed, is then stirred at room temperature 30min.4- (1- methyl isophthalic acid H- pyrazoles -4- bases) pyrimidine -2- amine (365mg, 2.081mmol) is added thereto, then at 75 DEG C Stir 16h.(TLC system 10%MeOH/ ethyl acetate, Rf:0.1).So that reactant mixture is cooled to room temperature and is diluted with water (100mL), is extracted with ethyl acetate (2x100mL).The organic layer of merging washs (50mL) with saturated brine solution, through anhydrous Na₂SO₄It is dried, filtered and concentrated, obtains crude compound.Crude product is through flash column chromatography (neutral alumina, eluant, eluent： 50%EtOAc/ petroleum ethers), obtain desired product (4S)-N- (4- (1- methyl isophthalic acid H- pyrazoles -4- bases) pyrimidine -2-base) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine (140mg, 0.307mmol, 22.11% yield), it is yellow solid.LCMS(m/z):454.19[M+H]⁺, Rt= 1.44min。

¹H NMR(400MHz,CDCl₃):δ ppm 13.75 (s, 1H), 8.59 (dd, J=12.06,5.26Hz, 2H), 7.95 (s, 1H), 7.90-7.84 (m, 2H), 7.76 (dd, J=5.15,1.43Hz, 1H), 7.64 (d, J=7.89Hz, 1H), 7.45 (d, J=7.89Hz, 1H), 7.08 (d, J=5.26Hz, 1H), 5.78 (dd, J=5.92,3.07Hz, 1H), 3.95 (s, 3H), 3.34-3.13 (m, 3H), 3.02 (dd, J=12.06,3.29Hz, 1H), 2.61 (s, 3H), 2.43-2.26 (m, 1H), 2.11 (dt, J=14.03,7.02Hz, 1H).

Embodiment 347

Synthesize (4S)-N- (1- methyl -5- (2- Jia Ji oxazole -5- bases) -2- oxo -1,2- dihydropyridine -3- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine

To (4S)-N- (the bromo- 1- methyl -2- oxos -1,2- dihydropyridines -3- bases of 5-) -7- (2- picoline -4- bases) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (700mg, 1.454mmol) (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- base) oxazoles (456mg, 2.181mmol) exist with 2- methyl -5- K is added in 1,4- dioxanes (24mL) and the solution of the stirring in water (6mL)₃PO₄(926mg,4.36mmol).Gained is reacted Mixture nitrogen degassing 15min.Then by PdCl₂(dppf) (106mg, 0.145mmol) adds to reactant mixture and again Deaerate 5min.Gained reactant mixture is stirred into 18h at 100 DEG C.(TLC systems:10%MeOH/DCM, R_f:0.6).Will reaction Mixture is diluted with water (30mL) and extracted (3x30mL) with EtOAc.The organic layer of merging water (20mL), saline solution (20mL) is washed, through anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain crude compound.Thick material is purified through column chromatography (silica gel:100-200 mesh；Eluant, eluent：4%MeOH/DCM), sticky solid is obtained.By the sticky solid with EtOH (1mL) With ether (15mL) washing, filter and fully dry, obtain desired product (4S)-N- (1- methyl -5- (2- Jia Ji oxazoles -5- Base) -2- oxo -1,2- dihydropyridine -3- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (95mg, 0.188mmol, 12.94% yield), it is pale solid. LCMS(m/z):484.24[M+H]⁺,R_t=1.51min.

¹H NMR(400MHz,CDCl₃):δ 12.46 (s, 1H), 8.61-8.53 (m, 2H), 8.09 (dt, J=1.9, 0.8Hz, 1H), 7.69-7.65. (d, J=5.6Hz, 1H), 7.61-7.59 (d, J=7.9Hz, 1H), 7.43-7.31 (m, 2H), 7.07 (s, 1H), 5.70 (dd, J=6.0,3.2Hz, 1H), 3.64 (s, 3H), 3.35-3.13 (m, 3H), 3.00 (dd, J= 12.1,3.3Hz, 1H), 2.69 (s, 3H), 2.49 (s, 3H), 2.39-2.34 (dddd, J=13.9,10.0,6.0,4.0Hz, 1H), 2.16-2.05 (dt, J=14.2,7.3Hz, 1H).

Embodiment 348

Synthesize (4S)-N- propyl group -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza- 5 (2H)-formamide

Room temperature to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diazaTriethylamine is added in the solution of (200mg, 0.655mmol) in tetrahydrofuran (20mL) (0.548mL, 3.93mmol) and triphosgene (194mg, 0.655mmol) and stirring 30min.Then add propyl- 1- amine (77mg, 1.310mmol) and at 80 DEG C heat 15h.(TLC systems:5%MeOH/DCM Rf:0.5).Reactant mixture is cooled to room Temperature, is concentrated in vacuo and by residue distribution between water (25mL) and EtOAc (70mL).Separate organic layer and through anhydrous sodium sulfate Dry, filter and filtrate evaporation is obtained into crude compound.Thick material is through flash column chromatography (100-200 silica gel, elution Agent：3% ethanol/methylene), obtain desired product (4S)-N- propyl group -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(130mg, 0.322mmol, 49.1% are received -5 (2H)-formamides Rate), it is pale solid.LCMS(m/z):391.07[M+H]⁺, Rt=2.47min.

¹H NMR(400MHz,CDCl₃):δ ppm 10.67-10.20 (m, 1H), 8.04 (s, 1H), 7.94 (d, J= 7.67Hz, 1H), 7.69-7.65 (m, 1H), 7.62-7.52 (m, 2H), 7.27 (s, 1H), 5.65 (dd, J=5.92,3.29Hz, 1H), 3.41 (m, 2H), 3.30 (m, 3H), 2.95 (dd, J=11.84,3.29Hz, 1H), 2.45 (m, 1H), 2.09 (m, 1H), 1.76-1.74 (m, 2H), 0.94 (t, J=7.45Hz, 3H).

Embodiment 349

Synthesize (4R) -7- (2- picoline -4- bases)-N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides

Room temperature to (4R) -7- chloro- N- (pyridin-3-yl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (3g, 9.50mmol) are at 1,4- dioxanes (80mL)：Stirring and degassing in water (20mL) Solution in add (2- picoline -4- bases) boric acid (1.952g, 14.25mmol) and K₃PO₄(6.05g,28.5mmol).So Reactant mixture is stirred into 15min afterwards.And argon-degassed 15min is used again.It is subsequently added into x-phos (1.812g, 3.80mmol) And Pd₂(dba)₃(1.740g, 1.900mmol), then stirs 3h at 110 DEG C.(TLC systems：Net EtOAc, Rf:0.2).Then So that reactant mixture is cooled to room temperature, frozen water (100mL) is poured into, is extracted with EtOAc (2 × 300mL).The organic layer warp of merging Anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain crude product.Thick material is through flash column chromatography (silica gel:100-200 Mesh, eluant, eluent：2%MeOH/DCM), obtain desired product (4R) -7- (2- picoline -4- bases)-N- (pyridin-3-yl) - 3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (2g, 5.37mmol, 56.5% yield), it is pale solid.LCMS(m/z):373.07[M+H]⁺,R_t=1.14min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.07 (s, 1H), 8.70-8.62 (m, 2H), 8.32 (dd, J=4.71, 1.43Hz, 1H), 8.20-8.09 (m, 1H), 7.65 (d, J=7.89Hz, 1H), 7.59 (s, 1H), 7.50 (dd, J=5.26, 1.32Hz, 1H), 7.38 (d, J=7.89Hz, 1H), 7.30-7.23 (m, 1H), 5.70 (dd, J=5.92,3.29Hz, 1H), 3.34-3.16 (m, 3H), 3.03 (dd, J=12.06,3.29Hz, 1H), 2.68 (s, 3H), 2.35 (dddd, J=14.14, 9.92,5.86,4.06Hz,1H),2.15-2.05(m,1H)。

Embodiment 350

Synthesis (4S)-N- cyclopenta -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza- 5 (2H)-formamides:

Room temperature to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diazaTEA is added in the solution of the stirring of (300.0mg, 0.983mmol) in THF (20mL) (0.685mL, 4.91mmol), triphosgene (292mg, 0.983mmol) simultaneously stir 45min.Then cyclopenta amine is added (0.291mL, 2.95mmol) and reactant mixture is heated to 75 DEG C, keeps 9h.(TLC elution systems:100%EtOAc；R_f- 0.4；UV activation).Reactant mixture is cooled to room temperature, solid is filtered and filtrate water is washed into (5mL) and is extracted into In EtOAc (2x15mL).Organic layer is separated, through anhydrous sodium sulfate drying, filters and filtrate evaporation is obtained into crude compound.Will Thick material is through chromatogram purification (neutral alumina, eluant, eluent：20-30% ethyl acetate/hexanes), obtain (4S)-N- cyclopenta -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formyls Amine (138.4mg, 0.331mmol, 33.7% yield), it is faint yellow solid.LCMS(m/z):417.24[M+H]⁺,R_t= 2.66min。

¹H NMR(400MHz,CDCl₃):δ ppm 10.37 (br d, J=6.14Hz, 1H), 8.02 (s, 1H), 7.94 (d, J =7.89Hz, 1H), 7.70-7.65 (m, 1H), 7.62-7.52 (m, 2H), 7.23 (d, J=7.89Hz, 1H), 5.66 (dd, J= 6.03,3.18Hz, 1H), 4.23 (sxt, J=6.88Hz, 1H), 3.30-3.06 (m, 3H), 2.94 (dd, J=12.06, 3.29Hz, 1H), 2.26 (dddd, J=13.98,9.98,5.97,4.06Hz, 1H), 2.10-1.99 (m, 3H), 1.59-1.72 (m,4H),1.56-1.44(m,2H)。

Embodiment 351

GSK3493927A

Synthesize (4S)-N- (3- hydroxypropyls) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C in a nitrogen atmosphere to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diazaAdded in the suspension of (500mg, 1.638mmol) in tetrahydrofuran (20mL) Simultaneously 1h is stirred at room temperature in reactant mixture by TEA (1.370mL, 9.83mmol) and triphosgene (486mg, 1.638mmol).In room Temperature adds solution of the 3- aminopropan-1-ols (246mg, 3.28mmol) in THF (5mL) to the obtained reactant mixture. Reactant mixture is stirred into 16h at 70 DEG C after addition.(TLC:5%MeOH-DCM, Rf value：0.5).Reactant mixture is cooled to RT is simultaneously diluted with water (50mL), is extracted with EtOAc (3x30mL).The organic layer of merging aqueous salt solu-tion (2x30mL), warp Anhydrous Na₂SO₄It is dried, filtered and concentrated, obtains crude compound.Crude product is through flash column chromatography (silica gel:100-200 Mesh, eluant, eluent：2%MeOH-DCM), desired product (4S)-N- (3- hydroxypropyls) -7- (3- (trifluoromethyl) benzene is obtained Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (180mg, 0.437mmol, 26.7% yield), it is pale solid.LCMS(m/z):407.09[M+H]⁺, Rt=1.95min.

¹H NMR(400MHz,DMSO-d₆):δ10.38-10.13(m,1H),8.23-8.13(m,2H),7.88-7.68(m, 2H), 7.66-7.54 (m, 2H), 5.41 (dd, J=5.81,2.96Hz, 1H), 4.43 (t, J=5.15Hz, 1H), 3.51-3.31 (m, 4H), 3.20-3.02 (m, 2H), 2.99-2.87 (m, 2H), 2.18 (dddd, J=13.73,9.95,5.92,3.73Hz, 1H), 1.82 (dt, J=13.98,7.15Hz, 1H), 1.69 (quin, J=6.69Hz, 2H).

Embodiment 352

Synthesis (4S)-N- cyclohexyl -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza- 5 (2H)-formamides:

Room temperature to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diazaTEA is added in the solution of the stirring of (300.0mg, 0.983mmol) in THF (20.0mL) (0.685mL, 4.91mmol), triphosgene (292mg, 0.983mmol) simultaneously stir 45min.Then cyclo-hexylamine is added (0.339mL, 2.95mmol) and reactant mixture is heated to 75 DEG C, keeps 6h.(TLC elution systems:100%EtOAc；R_f- 0.4；UV activation).Reactant mixture is cooled to room temperature, solid is filtered and filtrate water is washed into (5mL) and is extracted into In EtOAc (2 × 10mL).Organic layer is separated, through anhydrous sodium sulfate drying, filters and filtrate evaporation is obtained into crude compound. By thick material through chromatogram purification (neutral alumina, eluant, eluent：20-30% ethyl acetate/hexanes), obtain (4S)-N- cyclohexyl- 7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first Acid amides (221.9mg, 0.515mmol, 52.4% yield), it is faint yellow solid.LCMS(m/z):431.25[M+H]⁺,R_t= 2.82min。

¹H NMR(400MHz,CDCl₃):δ ppm 10.32 (br d, J=7.23Hz, 1H), 8.05 (s, 1H), 7.94 (d, J =7.89Hz, 1H), 7.70-7.66 (m, 1H), 7.62-7.52 (m, 2H), 7.28-7.25 (m, 1H), 5.65 (dd, J=5.92, 3.07Hz, 1H), 3.80-3.72 (m, 1H), 3.30-3.06 (m, 3H), 2.94 (dd, J=12.06,3.29Hz, 1H), 2.40- 2.26(m,1H),2.09-1.98(m,3H),1.77-1.62(m,3H),1.44-1.31(m,3H),1.29-1.10(m,2H)。

Embodiment 353

Synthesize (4S)-N- (tert-butyl group) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaIn the solution of (0.4g, 1.310mmol) in tetrahydrofuran (10mL) add triethylamine (1.096mL, 7.86mmol), triphosgene (0.389g, 1.310mmol), 1h is stirred at room temperature by reactant mixture, then adds 2- methyl propyl-s 2- amine (0.275mL, 2.62mmol), 16h is stirred by reactant mixture at 80 DEG C.(TLC systems：Net EtOAc.Rf values：0.5), Reactant mixture is cooled to room temperature, is extracted with ethyl acetate (2x30mL).The organic layer aqueous salt solu-tion of merging (2x20mL) and through anhydrous Na₂SO₄It is dried, filtered and concentrated, obtains crude compound.Crude product is through flash column chromatography (silicon Glue:100-200 mesh, eluant, eluent：5%MeOH/CH2Cl2), desired product (4S)-N- (tert-butyl group) -7- (3- (trifluoros are obtained Methyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (180mg, 0.436mmol, 33.3% yield), it is yellow solid.LCMS(m/z):405.11[M+H]⁺, Rt=2.67min.

¹H NMR(400MHz,DMSO-d₆):δppm 10.17(s,1H),8.08-8.16(m,2H),7.70-7.83(m, 2H), 7.60 (d, J=7.89Hz, 1H), 7.50 (d, J=7.89Hz, 1H), 5.41 (dd, J=5.81,2.96Hz, 1H), 3.01-3.16(m,2H),2.86-2.97(m,2H),2.12-2.21(m,1H),1.75-1.85(m,1H),1.28(s,9H)。

Embodiment 354

Synthesize (4S)-N- (4- (2- Jia Ji oxazole -5- bases) pyrimidine -2-base) -7- (2- picoline -4- bases) -3,4- two Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4S)-N- (4- chlorine pyrimidine -2-base) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (700mg, 1.716mmol) are in 1,4- dioxanes (25mL) and water In (5.00mL) PdCl is added in the solution of degassing₂(dppf)-CH₂Cl₂Adduct (70.1mg, 0.086mmol), K₃PO₄ (1093mg, 5.15mmol) and 2- methyl -5- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- base) oxazoles (466mg, 2.231mmol) and stir 3h at 70 DEG C.(TLC systems:10% methanol/DCM, Rf values：0.4).To reactant mixture Middle addition water (10.0mL), and (3 × 30mL) is extracted with ethyl acetate.The separation organic layer of merging, through anhydrous Na₂SO₄Dry, Filter and filtrate evaporation is obtained into crude product.Crude compound purifies (post through preparation HPLC：KROMASIL PHENYL(150* 25)mm*10u；MP-A:10mM ammonium hydrogen carbonate (aqueous solution), MP-B：Acetonitrile；Method：00/30,13.5/30,14/100,18/ 100,18.5/30；Flow velocity：25ml/min；Temperature:Environment temperature；Eluant, eluent：Acetonitrile+THF+ water), obtain (4S)-N- (4- (2- Jia Ji oxazole -5- bases) pyrimidine -2-base) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza- 5 (2H)-formamides (120mg, 0.258mmol, 15.04% yield), it is faint yellow solid.LCMS (m/z):455.19[M+H]⁺, Rt=1.35min.

¹H NMR(400MHz,CDCl₃):δppm:13.87 (s, 1H), 8.69 (d, J=5.04Hz, 1H), 8.63 (d, J= 5.26Hz, 1H), 7.89 (s, 1H), 7.81-7.76 (m, 1H), 7.71 (s, 1H), 7.64 (d, J=7.89Hz, 1H), 7.48 (d, J=7.89Hz, 1H), 7.23 (d, J=5.04Hz, 1H), 5.79 (dd, J=5.92,3.07Hz, 1H), 3.33-3.13 (m, 3H), 3.02 (dd, J=12.17,3.18Hz, 1H), 2.65 (s, 3H), 2.59 (s, 3H), 2.39-2.30 (m, 1H), 2.11 (dt, J=14.47,7.45Hz, 1H).

Embodiment 355

Synthesize (4S) -7- (2- picoline -4- bases)-N- (4- morpholino pyridine -2- bases) -3,4- dihydro -1,4- bridges sub- Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

In a nitrogen atmosphere in room temperature to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diazaIn the solution of the stirring of (300mg, 1.189mmol) in tetrahydrofuran (30mL) Triethylamine (0.829mL, 5.94mmol) and triphosgene (353mg, 1.189mmol) are added, 1h is then stirred at room temperature.Then will 4- morpholino pyridine -2- amine (320mg, 1.783mmol) adds to reactant mixture, then stirs 16h at 70 DEG C.(TLC systems: 10%MeOH/DCM, R_f-0.4；UV activation).Reactant mixture is cooled to room temperature and distributed in water (30mL) and EtOAc Between (3X30mL).The organic layer aqueous salt solu-tion of merging, through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain Crude product.Crude compound purifies (silica gel through column chromatography:Neutral alumina, eluant, eluent：20%EtOAc/ petroleum ethers), it must expire Product (4S) -7- (2- picoline -4- bases)-N- (4- morpholino pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups of prestige Pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (145mg, 0.312mmol, 26.3% yield), it is greyish white Color solid.LCMS(m/z):458.29[M+H]⁺,R_t=1.23min.

¹H NMR(400MHz,CDCl₃):δ 13.40 (s, 1H), 8.61 (d, J=5.2Hz, 1H), 8.24 (d, J=1.8Hz, 1H), 8.09 (d, J=6.0Hz, 1H), 7.79 (d, J=2.4Hz, 1H), 7.75-7.68 (m, 1H), 7.61 (d, J=8.0Hz, 1H), 7.48 (d, J=7.9Hz, 1H), 6.45 (dd, J=6.0,2.5Hz, 1H), 5.68 (dd, J=6.0,3.2Hz, 1H), 3.88-3.80 (m, 4H), 3.40-3.32 (m, 4H), 3.32-3.20 (m, 2H), 3.24-3.13 (m, 1H), 3.01 (dd, J= 12.1,3.3Hz, 1H), 2.73 (s, 3H), 2.40-2.26 (m, 1H), 2.08 (dt, J=14.9,7.7Hz, 1H).

Embodiment 356

Synthesize (4S) -7- (2- picoline -4- bases)-N- (4- (oxazole -5- bases) pyrimidine -2-base) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4S)-N- (4- chlorine pyrimidine -2-base) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (600mg, 1.471mmol), 5- (4,4,5,5- tetramethyl -1,3, 2- dioxaborolan -2- base) oxazoles (430mg, 2.207mmol) and K₃PO₄(937mg, 4.41mmol) is in 1,4- dioxanes In (30.00mL), water (7.50mL) PdCl is added in the solution of degassing₂(dppf)-CH₂Cl₂Adduct (120mg, Deaerate 0.147mmol) and again 5min.Then reactant mixture is stirred into 15h at 80 DEG C.(TLC systems:10% methanol/DCM.R_f Value：0.2).So that reactant mixture is cooled to RT and evaporation of organic solvent, (50mL) is diluted with water and is extracted with ethyl acetate (3x50ml).The organic layer of merging is washed with saturated brine solution, then through anhydrous sodium sulfate drying, is filtered and is evaporated in vacuo, Obtain crude product.Crude compound is through flash column chromatography (neutral alumina, eluant, eluent：2% methanol/ethyl acetate), obtain To (4S) -7- (2- picoline -4- bases)-N- (4- (oxazole -5- bases) pyrimidine -2-base) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (320mg, 0.710mmol, 48.3% yield), it is canescence Solid.LCMS(m/z):441.23[M+H]⁺, Rt=1.30min.

¹H NMR(400MHz,CDCl₃):δ 13.91 (s, 1H), 8.74 (dd, J=5.1,0.6Hz, 1H), 8.64 (d, J= 5.2Hz, 1H), 8.05 (d, J=0.5Hz, 1H), 7.90-7.76 (m, 2H), 7.65 (dd, J=8.0,0.6Hz, 1H), 7.48 (dd, J=8.0,0.6Hz, 1H), 7.33-7.24 (m, 2H), 5.79 (dd, J=6.0,3.2Hz, 1H), 3.35-3.12 (m, 3H), 3.03 (dd, J=12.1,3.3Hz, 1H), 2.65 (s, 3H), 2.35 (dddd, J=14.1,9.9,6.0,4.0Hz, 1H), 2.11 (dt, J=14.6,7.4Hz, 1H).

Embodiment 357

Synthesize (4S)-N- (4- ((2S, 6R) -2,6- thebaine generations) pyridine -2- bases) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Under a nitrogen in room temperature to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diazaThe solution of the stirring of (300mg, 1.189mmol) in tetrahydrofuran (THF) (30mL) Middle addition triethylamine (0.829mL, 5.94mmol) and triphosgene (353mg, 1.189mmol), are then stirred at room temperature 4h.Then 4- ((2S, 6R) -2,6- thebaine generations) pyridine -2- amine (493mg, 2.378mmol) is added into reactant mixture, by gained Reactant mixture stirs 16h at 70 DEG C.(TLC systems:10%MeOH/DCM, R_f-0.4；UV activation).Reactant mixture is cold But to room temperature, distribute between water (30mL) and EtOAc (3X30mL).The organic layer aqueous salt solu-tion of merging, through anhydrous Na₂SO₄Dry, filter and evaporate filtrate, obtain crude compound.Crude compound is through flash column chromatography (silica gel:In Property aluminum oxide, eluant, eluent：25%EtOAc/ petroleum ethers), obtain desired product (4S)-N- (4- ((2S, 6R) -2,6- dimethyl Morpholino) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] Diaza- 5 (2H)-formamides (200mg, 0.410mmol, 34.5% yield), it is pale solid.LCMS(m/z): 486.33[M+H]⁺,R_t=1.37min.

¹H NMR(400MHz,CDCl₃):δ 13.40 (s, 1H), 8.60 (d, J=5.3Hz, 1H), 8.25 (d, J=1.8Hz, 1H), 8.07 (d, J=6.0Hz, 1H), 7.78-7.68 (m, 2H), 7.61 (d, J=7.9Hz, 1H), 7.48 (d, J=8.0Hz, 1H), 6.45 (dd, J=6.0,2.5Hz, 1H), 5.69 (dd, J=6.0,3.2Hz, 1H), 3.81-3.64 (m, 4H), 3.32- 3.12 (m, 3H), 3.01 (dd, J=12.1,3.3Hz, 1H), 2.73 (s, 3H), 2.61-2.51 (m, 2H), 2.34 (dddd, J= 13.9,10.0,6.0,4.0Hz, 1H), 2.09 (dt, J=14.3,7.8Hz, 1H), 1.28 (d, J=6.2Hz, 6H).

Embodiment 358

Synthesize (4S)-N- (4- (1,3- dimethyl -1H- pyrazoles -4- bases) pyrimidine -2-base) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4S)-N- (4- chlorine pyrimidine -2-base) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (200mg, 0.490mmol), 1,3- dimethyl -4- (4,4,5,5- Tetramethyl -1,3,2- dioxaborolan -2- bases) -1H- pyrazoles (109mg, 0.490mmol) and K₃PO₄(104mg, 0.490mmol) PdCl is added in 1,4- dioxanes (30.00mL), water (7.50mL) in the solution of degassing₂(dppf)-CH₂Cl₂ Adduct (400mg, 0.490mmol) and the 5min that deaerates again.Then reactant mixture is stirred into 15h at 80 DEG C.(TLC systems: 10% methanol/DCM.R_fValue：0.2).Reactant mixture is cooled to room temperature and evaporation of organic solvent, (50mL) is diluted with water simultaneously It is extracted with ethyl acetate (3x50ml).The organic layer of merging is washed with saturated brine solution, then through anhydrous sodium sulfate drying simultaneously It is evaporated in vacuo, obtains crude product.Crude compound is through flash column chromatography (neutral alumina, eluant, eluent：4% methanol/acetic acid Ethyl ester), obtain desired product (4S)-N- (4- (1,3- dimethyl -1H- pyrazoles -4- bases) pyrimidine -2-base) -7- (2- methyl pyrroles Pyridine -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (120mg, 0.256mmol, 52.2% yield), it is pale solid.LCMS(m/z):468.30[M+H]⁺, Rt=1.36min.

¹H NMR(400MHz,CDCl₃):δ 13.68 (s, 1H), 8.58 (dd, J=6.6,5.3Hz, 1H), 7.95 (s, 1H), 7.85 (s, 1H), 7.79 (s, 1H), 7.65 (d, J=8.0Hz, 1H), 7.46 (d, J=7.9Hz, 1H), 7.26 (s, 1H), 7.09 (d, J=5.2Hz, 1H), 5.79 (dd, J=6.0,3.1Hz, 1H), 3.88 (s, 3H), 3.32-3.12 (m, 3H), 3.02 (dd, J =12.0,3.3Hz, 1H), 2.64 (s, 3H), 2.54 (s, 3H), 2.35 (ddt, J=14.3,10.0,4.7Hz, 1H), 2.11 (dt, J=14.8,7.8Hz, 1H).

Embodiment 359,360

Peak-I:(4S)-N- (4- (3,4- dihydroxy butyl) pyridine -2- bases) -7- (2- picoline -4- bases) -3 is synthesized, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Peak-II:(4S)-N- (4- (3,4- dihydroxy butyl) pyridine -2- bases) -7- (2- picoline -4- bases) -3 is synthesized, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (4S)-N- (4- (2- (1,4- dioxo spiros [4.5] decyl- 2- yls) ethyl) pyridine -2- bases) -7- (2- Picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides HCl (2mL, 24.00mmol) is added in the solution of the stirring of (850mg, 1.572mmol) in methanol (10mL).Gained is anti- Answer mixture that 6h is stirred at room temperature.(TLC systems:10%MeOH/DCM, R_f:And evaporation solvent, 0.5) residue of acquisition is used NaHCO₃Solution is neutralized and extracted (3 × 30mL) with DCM.The organic layer of merging is with aqueous salt solu-tion (20mL) and through anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain crude compound.Crude compound is through flash column chromatography (silica gel:100- 200 mesh, eluant, eluent：6%MeOH/DCM), the chiral SFC of racemic compound (400mg) .The racemic compounds is obtained pure (the condition of change:Post/size:LuxAmylose-2 (250X30) mm, 5 μ, %CO2 solvents：50.0%, %CO solvent：50.0% (100%IPA), overall flow rate：90.0g/min, back-pressure:100.0 bar, UV:218nm, accumulation time (Stack time): 7.5min, load/injection:20.0mg, eluant, eluent：IPA+DCM, instrument details:Preparation/model:Thar SFC-200(NEW- 2)) peak-I:(4S)-N- (4- (3,4- dihydroxy butyl) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(163mg, 0.353mmol, 22.48% are received -5 (2H)-formamides Rate), it is pale solid.LCMS(m/z):461.35[M+H]⁺, Rt=1.25min, and peak-II:(4S)-N-(4-(3,4- Dihydroxy butyl) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (67mg, 0.142mmol, 9.03% yield), it is the solid of clear yellow viscous.LCMS (m/z):461.31[M+H]⁺, Rt=1.25min.

Peak-I:¹H NMR(400MHz,CDCl₃):δ 13.52 (s, 1H), 8.61 (d, J=5.3Hz, 1H), 8.26 (dd, J= 5.0,0.8Hz, 1H), 8.20 (d, J=1.8Hz, 1H), 8.13 (s, 1H), 7.71 (dd, J=5.4,1.8Hz, 1H), 7.62 (d, J=8.0Hz, 1H), 7.48 (d, J=8.0Hz, 1H), 6.89 (dd, J=5.1,1.5Hz, 1H), 5.69 (dd, J=6.0, 3.2Hz, 1H), 3.81-3.64 (m, 2H), 3.49 (dd, J=11.0,7.4Hz, 1H), 3.35-3.13 (m, 3H), 3.02 (dd, J =12.0,3.3Hz, 1H), 2.92-2.74 (m, 2H), 2.73 (s, 3H), 2.34 (ddt, J=14.5,10.0,5.1Hz, 1H), 2.09 (dt, J=14.5,7.4Hz, 1H), 1.88-1.77 (m, 2H).

Peak-II:¹H NMR(400MHz,CDCl₃):δ 13.52 (s, 1H), 8.61 (d, J=5.3Hz, 1H), 8.26 (dd, J =5.0,0.8Hz, 1H), 8.20 (d, J=1.8Hz, 1H), 8.13 (s, 1H), 7.71 (dd, J=5.4,1.8Hz, 1H), 7.62 (d, J=8.0Hz, 1H), 7.48 (d, J=8.0Hz, 1H), 6.89 (dd, J=5.1,1.5Hz, 1H), 5.69 (dd, J=6.0, 3.2Hz, 1H), 3.81-3.64 (m, 2H), 3.49 (dd, J=11.0,7.4Hz, 1H), 3.35-3.13 (m, 3H), 3.02 (dd, J =12.0,3.3Hz, 1H), 2.92-2.74 (m, 2H), 2.73 (s, 3H), 2.34 (ddt, J=14.5,10.0,5.1Hz, 1H), 2.09 (dt, J=14.5,7.4Hz, 1H), 1.88-1.77 (m, 2H).

Embodiment 361

Synthesize (4S) -7- (6- (methylamino) pyridin-3-yl)-N- (pyridine -2- bases) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

To the chloro- N- of (4S) -7- (pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] phenodiazine It is miscellaneousAdd in the solution of stirring of -5 (the 2H)-formamides (3g, 9.50mmol) in 1,4- dioxanes (70mL) and water (24mL) Enter N- methyl -5- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) pyridine -2- amine (2.447g, 10.45mmol) and K₃PO₄(6.05g, 28.5mmol) and the 20min that deaerates.Then by Pd₂(dba)₃(0.435g,0.475mmol) Reactant mixture is added to X-phos (0.453g, 0.950mmol) and the 10min that deaerates again.Reactant mixture is stirred at 100 DEG C Mix 16h.(TLC eluant, eluents：Net ethyl acetate:R_f-0.4；UV activation).Reactant mixture is cooled to room temperature and dilute with water Release, be extracted with ethyl acetate (2X70mL), the organic layer of merging is with aqueous salt solu-tion (50mL), through anhydrous sodium sulfate drying, Filter and concentrate, obtain crude compound.Crude product (uses 100-200 silica gel, compound is 70% through flash column chromatography In ethyl acetate/n-hexane elute), obtain desired product (4S) -7- (6- (methylamino) pyridin-3-yl)-N- (pyridine - 2- yls) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (1.47g, 3.78mmol, 39.8% yield), it is white solid.LCMS(m/z):388.13[M+H]⁺,R_t=1.38min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.69 (s, 1H), 8.75 (d, J=2.19Hz, 1H), 8.52 (dd, J= 8.77,2.63Hz, 1H), 8.43-8.29 (m, 1H), 8.29-8.06 (m, 1H), 7.68 (t, J=7.84Hz, 1H), 7.52 (d, J =7.89Hz, 1H), 7.33-7.19 (m, 1H), 6.97 (ddd, J=7.23,4.82,0.88Hz, 1H), 6.55 (d, J= 8.99Hz, 1H), 5.68 (dd, J=5.92,3.07Hz, 1H), 4.77 (d, J=4.60Hz, 1H), 3.33-3.08 (m, 3H), 3.06-2.94 (m, 4H), 2.31 (dddd, J=14.03,9.92,5.97,3.84Hz, 1H), 2.17-1.93 (m, 1H).

Embodiment 362

Synthesize (4S)-N- (5- Fluquinconazole quinoline -4- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- bridge methylenes Yl pyridines simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides:

Add in the solution of 0 DEG C of stirring to 5- Fluquinconazole quinoline -4- amine (250mg, 1.532mmol) in THF (10mL) Enter NaH (201mg, 4.60mmol) and stir 10min.Then (1H- imidazoles -1- bases) ((4S) -7- (3- (trifluoromethyl) are added Phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-yls) ketone (612mg, 1.532mmol) and by reactant mixture at 60 DEG C stir 12h. (TLC elution systems:10%MeOH/EtOAc；R_f-0.3；UV lives Change).Reactant mixture is cooled to room temperature, (10mL) is quenched with water and is extracted into EtOAc (25mL).Separate organic layer and Through anhydrous sodium sulfate drying, filter and filtrate evaporation is obtained into crude compound.By thick material, through chromatogram purification, (GRACE is used C-18 reversed-phase columns, mobile phase A:0.1% formic acid/water；B:MeOH, eluant, eluent 65%B/A).The fraction of merging is concentrated, and with satisfying And NaHCO₃It is basified.Water layer is extracted with DCM, the DCM layers isolated are through anhydrous Na₂SO₄Dry, filter and steam filtrate Hair, obtains (4S)-N- (5- Fluquinconazole quinoline -4- bases) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro-Isosorbide-5-Nitrae-endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (190mg, 0.380mmol, 24.77% yield), it is solid for white Body.LCMS(m/z):495.08[M+H]⁺,R_t=2.19min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.86 (br d, J=4.60Hz, 1H), 9.13 (s, 1H), 7.95 (s, 1H), 7.76-7.88 (m, 2H), 7.68-7.75 (m, 1H), 7.66 (d, J=7.89Hz, 1H), 7.60 (d, J=7.89Hz, 1H), 7.32-7.42 (m, 2H), 6.75-6.84 (m, 1H), 5.78 (dd, J=5.92,3.29Hz, 1H), 3.17-3.38 (m, 3H), 3.05 (dd, J=12.28,3.29Hz, 1H), 2.31-2.43 (m, 1H), 2.13-2.24 (m, 1H).

Embodiment 363

Synthesize (4S)-N- methyl -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza- 5 (2H)-formamides

Room temperature to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diazaTriethylamine is added in the solution of the stirring of (500mg, 1.638mmol) in THF (25mL) (1.370mL, 9.83mmol), is subsequently added into triphosgene (486mg, 1.638mmol) and stirs 1h.Then 2M methylamines are added Reactant mixture is simultaneously heated 16h by THF solution (2.457mL, 4.91mmol) at 65 DEG C.(TLC eluant, eluents：50%EtOAc/ oneself Alkane:R_f-0.3；UV activation).Reactant mixture is cooled to room temperature, be concentrated in vacuo and by residue distribution in water (30mL) and Between EtOAc (2x35mL).Separate organic layer and through anhydrous sodium sulfate drying, filter and filtrate evaporation is obtained into rough chemical combination Thing.Crude product is through flash column chromatography (neutral alumina, eluant, eluent：50% ethyl acetate/hexane), obtain (4S)-N- first Base -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (161mg, 0.444mmol, 27.1% yield), it is white solid.LCMS(m/z):362.99[M+H]⁺,R_t =2.19min.

¹H NMR(400MHz,CDCl₃):δ ppm 10.38 (br d, J=3.51Hz, 1H), 8.06 (s, 1H), 7.94 (d, J =7.67Hz, 1H), 7.70-7.66 (m, 1H), 7.63-7.58 (m, 1H), 7.55 (d, J=7.89Hz, 1H), 7.28 (d, J= 7.89Hz, 1H), 5.65 (dd, J=6.03,3.18Hz, 1H), 3.28-3.07 (m, 3H), 3.01-2.92 (m, 4H), 2.27 (dddd, J=13.98,9.92,6.03,4.06Hz, 1H), 2.08-1.97 (m, 1H).

Embodiment 364

Synthesize (4S)-N- ((R) -2,3- dihydroxypropyls) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides, hydrochloride

(4S)-N- (4- (3- methyl isophthalic acid H- pyrazoles -4- bases) pyridine -2- bases) -7- (2- picoline -4- bases) -3 is synthesized, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

It is sub- to (4S)-N- (4- bromopyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridges in room temperature Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (300mg, 0.665mmol) are in 1,4- dioxanes 3- methyl -4- is added in (40mL) and the solution of the stirring in water (10mL), and (4,4,5,5- tetramethyl -1,3,2- dioxa boron is miscellaneous The amyl- 2- yls of ring) -1H- pyrazoles (207mg, 0.997mmol) and K₃PO₄(423mg, 1.994mmol) and the 15min that deaerates.Then will Pd₂(dba)₃(60.9mg, 0.066mmol) and x-phos (63.4mg, 0.133mmol) add to reactant mixture, deaerate again 10min, then stirs 16h by reactant mixture at 80 DEG C.(TLC systems:10% methanol/DCM, R_f-0.2；UV activation).Will Reactant mixture is cooled to room temperature and distributed between water and ethyl acetate (3x30mL).The organic layer of merging is washed with saline solution Wash, through anhydrous Na₂SO₄Dry and be concentrated under reduced pressure, obtain crude compound.Crude compound is through flash column chromatography (in use Property aluminum oxide, in 20% ethyl acetate/petroleum ether elute), obtain desired compound (4S)-N- (4- (3- methyl isophthalic acid H- pyrroles Azoles -4- bases) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (139mg, 0.304mmol, 45.7% yield), it is Light brown solid.LCMS(m/z): 453.21[M+H]⁺,R_t=1.36min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.59 (s, 1H), 10.35 (br s, 1H), 8.63 (d, J=5.26Hz, 1H), 8.44-8.16 (m, 3H), 7.87 (s, 1H), 7.73 (d, J=3.95Hz, 1H), 7.63 (d, J=8.11Hz, 1H), 7.49 (d, J=7.89Hz, 1H), 7.10 (dd, J=5.15,1.43Hz, 1H), 5.72 (dd, J=5.70,3.07Hz, 1H), 3.36- 3.16 (m, 3H), 3.02 (dd, J=12.06,3.07Hz, 1H), 2.76 (s, 3H), 2.59 (s, 3H), 2.42-2.29 (m, 1H), 2.10 (dt, J=14.03,7.02Hz, 1H).

Embodiment 365

Synthesize (4S) -7- (2- picoline -4- bases)-N- (4- (2- oxo-pyrrolidine -1- bases) pyridine -2- bases) -3,4- Dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In a nitrogen atmosphere to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diazaTriethylamine is added in the solution of the stirring of (400mg, 1.585mmol) in THF (40mL) (1.105mL, 7.93mmol) and triphosgene (470mg, 1.585mmol), is then stirred at room temperature 1h.1- (2- are added thereto Aminopyridine -4- bases) pyrrolidin-2-one (562mg, 3.17mmol), then stirs 16h at 70 DEG C.(TLC eluant, eluents：10% MeOH/DCM,R_f-:0.3, UV activation).Reactant mixture is cooled to room temperature, distributed between water and ethyl acetate (3X40mL).The organic layer aqueous salt solu-tion of merging, through anhydrous Na₂SO₄Dry, then evaporation obtains crude compound.Slightly Produced compounds are expected through flash column chromatography (using neutral alumina, eluted in 25% ethyl acetate/petroleum ether) Compound (4S) -7- (2- picoline -4- bases)-N- (4- (2- oxo-pyrrolidine -1- bases) pyridine -2- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza(287mg, 0.625mmol, 39.5% are received -5 (2H)-formamides Rate), it is pale solid.LCMS(m/z):456.20[M+H]⁺,R_t=1.40min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.59 (s, 1H), 8.62 (d, J=5.26Hz, 1H), 8.29 (d, J= 5.92Hz, 1H), 8.21 (s, 1H), 8.12 (d, J=1.97Hz, 1H), 7.92 (dd, J=5.70,2.19Hz, 1H), 7.71 (dd, J=5.26,1.53Hz, 1H), 7.62 (d, J=7.89Hz, 1H), 7.48 (d, J=8.11Hz, 1H), 5.69 (dd, J= 6.03,3.18Hz, 1H), 3.96 (t, J=7.13Hz, 2H), 3.35-3.13 (m, 3H), 3.02 (dd, J=12.06,3.29Hz, 1H), 2.74 (s, 3H), 2.65 (t, J=8.11Hz, 2H), 2.40-2.28 (m, 1H), 2.19 (quin, J=7.62Hz, 2H), 2.13-2.02(m,1H)。

Embodiment 366

Synthesize (4S) -7- (2- picoline -4- bases)-N- (4- (1,3,5- trimethyl -1H- pyrazoles -4- bases) pyridine -2- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

It is sub- to (4S)-N- (4- bromopyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridges in room temperature Picoline simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (300mg, 0.665mmol) are in 1,4- dioxanes 1,3,5- trimethyls -4- (4,4,5,5- tetramethyl -1,3,2- dioxies are added in (30mL) and the solution of the stirring in water (6mL) The miscellaneous amyl- 2- yls of boron heterocycle) -1H- pyrazoles (235mg, 0.997mmol) and K₃PO₄(423mg, 1.994mmol), deaerate 10min.So Afterwards by PdCl₂(dppf)-CH₂Cl₂Adduct (54.3mg, 0.066mmol) adds to reactant mixture and the 5min that deaerates again.Will Reactant mixture is stirred to 80 DEG C, keeps 16h.(TLC eluant, eluents：10% methanol/DCM, Rf:0.3；UV activation).Will reaction Mixture is cooled to room temperature, distributes between water and ethyl acetate (3X30mL).The organic layer of merging is washed with saturated brine solution Wash and through anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain crude compound.Crude compound is through flash column chromatography (using neutral alumina, eluted in 30% ethyl acetate/petroleum ether), obtains pure compound (4S) -7- (2- methyl pyrroles Pyridine -4- bases)-N- (4- (1,3,5- trimethyl -1H- pyrazoles -4- bases) pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyridines And [2,3-b] [1,4] diaza- 5 (2H)-formamides (238mg, 0.494mmol, 74.2% yield), it is solid for canescence Body.LCMS(m/z):481.22[M+H]⁺,R_t=1.55min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.57 (s, 1H), 8.63 (d, J=5.26Hz, 1H), 8.37 (dd, J= 5.04,0.66Hz, 1H), 8.24 (s, 1H), 8.16 (d, J=0.66Hz, 1H), 7.73 (dd, J=5.26,1.53Hz, 1H), 7.62 (d, J=7.89Hz, 1H), 7.49 (d, J=7.89Hz, 1H), 6.93 (dd, J=5.26,1.53Hz, 1H), 5.71 (dd, J=5.92,3.29Hz, 1H), 3.79 (s, 3H) 3.36-3.13 (m, 3H), 3.01 (dd, J=12.06,3.29Hz, 1H), 2.76 (s, 3H), 2.36 (d, J=7.67Hz, 7H), 2.09 (dt, J=14.25,7.34Hz, 1H).

Embodiment 367

Synthesize (4S) -7- (2- picoline -4- bases)-N- (4- (1- (2,2,2- trifluoroethyls) -1H- pyrazoles -4- bases) pyrroles Pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Under argon gas to (4S)-N- (4- bromopyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (450mg, 0.997mmol) are in 1,4- dioxanes In (50mL) and water (10mL) K is added in the solution of degassing₃PO₄(635mg, 2.99mmol), 4- (4,4,5,5- tetramethyl -1,3, 2- dioxaborolan -2- bases) -1- (2,2,2- trifluoroethyl) -1H- pyrazoles (358mg, 1.296mmol), then in room temperature Stir 10min.Then PdCl is added₂(dppf)-CH₂Cl₂Adduct (81mg, 0.100mmol) and 80 DEG C stir 3h.(TLC System:10% methanol/DCM, Rf values：0.4).Water (100mL) is added into reactant mixture and is extracted with ethyl acetate (3x100mL).By the extract of merging with aqueous salt solu-tion (100mL), through anhydrous Na₂SO₄Dry, filter and steam filtrate Hair obtains crude compound.Crude product purifies (post through preparation HPLC：XBridge C-18(150*19)mm,5μ；Mobile phase- A:0.1% formic acid；Mobile phase-B：Acetonitrile；Flow velocity：15mL/min；Method：0.1/37,1/37,10/37；Eluant, eluent：Acetonitrile+ THF+MeOH), (4S) -7- (2- picoline -4- bases)-N- (4- (1- (2,2,2- trifluoroethyl) -1H- pyrazoles -4- bases) are obtained Pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (155mg, 0.297mmol, 29.8% yield), it is pale solid.LCMS(m/z):521.22[M+H]⁺, Rt= 1.74min。

¹H NMR(400MHz,CDCl₃):δ ppm 13.61 (s, 1H), 8.63 (d, J=5.48Hz, 1H), 8.41 (s, 1H), 8.35 (d, J=5.04Hz, 1H), 8.21 (s, 1H), 8.00 (s, 1H), 7.96 (s, 1H), 7.73 (br d, J=4.38Hz, 1H), 7.63 (d, J=7.89Hz, 1H), 7.49 (d, J=7.89Hz, 1H), 7.13 (d, J=3.95Hz, 1H), 5.71 (dd, J =5.70,3.07Hz, 1H), 4.75 (q, J=8.26Hz, 2H), 3.36-3.12 (m, 3H), 3.03 (dd, J=12.06, 3.07Hz, 1H), 2.75 (s, 3H), 2.36 (qd, J=9.83,4.49Hz, 1H), 2.11 (dt, J=14.20,7.26Hz, 1H).

Embodiment 368

(4S) -7- (2- picoline -4- bases)-N- (4- (pyrrolidin-1-yl) pyridine -2- bases) -3,4- dihydro -1 is synthesized, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Under a nitrogen to (4S) -7- (2- picoline -4- bases) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diaza(add triethylamine in the solution of the stirring of 300mg, 1.189mmol in THF (30mL) (0.829mL, 5.94mmol) and triphosgene (353mg, 1.189mmol), is stirred at room temperature 1h.Thereto add 4- (pyrrolidines- 1- yls) pyridine -2- amine (388mg, 2.378mmol), then stirs 16h at 70 DEG C.(TLC eluant, eluents：10%MeOH/DCM, R_f-:0.4, UV activation).Reactant mixture is cooled to room temperature, distributed between water and ethyl acetate (3X30mL).Merge Organic layer aqueous salt solu-tion, through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into crude compound.Roughization Compound obtains desired product through flash column chromatography (using neutral alumina, eluted in 20%EtOAc/ petroleum ethers) (4S) -7- (2- picoline -4- bases)-N- (4- (pyrrolidin-1-yl) pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (251mg, 0.561mmol, 47.2% yield), it is canescence Solid.LCMS(m/z):442.3[M+H]⁺,R_t=4.78min.

¹H NMR(400MHz,CDCl₃):δ 13.27 (s, 1H), 8.60 (d, J=5.3Hz, 1H), 8.28 (s, 1H), 8.00 (d, J=5.9Hz, 1H), 7.72 (dd, J=5.5,1.8Hz, 1H), 7.59 (d, J=7.9Hz, 1H), 7.50-7.38 (m, 2H), 6.19 (dd, J=5.9,2.3Hz, 1H), 5.70 (dd, J=6.0,3.2Hz, 1H), 3.39 (q, J=6.2,4.8Hz, 3H), 3.26 (s, 1H), 3.27-3.12 (m, 3H), 3.00 (dd, J=12.0,3.3Hz, 1H), 2.74 (s, 3H), 2.32 (dddd, J= 14.0,10.0,6.2,4.2Hz,1H),2.15-1.97(m,5H)。

Embodiment 369

Synthesize (4S)-N- (4- (1,3- dimethyl -1H- pyrazoles -4- bases) pyridine -2- bases) -9- methyl -7- (2- methyl pyrroles Pyridine -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (4S)-N- (4- bromopyridine -2- bases) -9- methyl -7- (2- picoline -4- bases) -3,4- dihydro -1, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (300mg, 0.645mmol) are in 1,4- bis- Evil 1,3- dimethyl -4- (4,4,5,5- tetramethyl -1,3,2- dioxies are added in alkane (40mL) and the solution of the stirring in water (10mL) The miscellaneous amyl- 2- yls of boron heterocycle) -1H- pyrazoles (215mg, 0.967mmol) and K₃PO₄(411mg, 1.934mmol) and the 15min that deaerates. Then PdCl is added₂(dppf)-CH₂Cl₂Adduct (52.6mg, 0.064mmol) and the 5min that deaerates, then in 80 DEG C of stirrings 16h.(TLC eluant, eluents：Net ethyl acetate:Rf:0.3；UV activation).Reactant mixture is cooled to room temperature and frozen water is poured into In, it is extracted with ethyl acetate (2x100ml).The organic layer of merging is washed with saturated brine (50mL) solution, then through anhydrous sulphur Sour sodium is dried and is evaporated in vacuo, and obtains crude product.Crude compound through flash column chromatography (use neutral alumina, Post is eluted in 60% ethyl acetate/petroleum ether), obtain pure compound (4S)-N- (4- (1,3- dimethyl -1H- pyrazoles -4- Base) pyridine -2- bases) -9- methyl -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] diaza- 5 (2H)-formamides (101mg, 0.208mmol, 32.3% yield), it is pale solid.LCMS(m/ z):481.33[M+H]⁺,R_t=1.70min.

¹H NMR(400MHz,CDCl₃):δ 13.72 (s, 1H), 8.61 (d, J=5.3Hz, 1H), 8.36-8.29 (m, 2H), 8.22 (d, J=1.7Hz, 1H), 7.76-7.70 (m, 1H), 7.67 (s, 1H), 7.38 (d, J=0.7Hz, 1H), 7.06 (dd, J =5.2,1.7Hz, 1H), 5.71 (dd, J=6.1,3.1Hz, 1H), 3.88 (s, 3H), 3.22-3.07 (m, 3H), 3.02 (dd, J =12.0,3.2Hz, 1H), 2.75 (s, 3H), 2.51 (d, J=2.8Hz, 6H), 2.33 (m, 1H), 2.13-2.00 (m, 1H).

Embodiment 370

Synthesize (4S) -9- methyl-N- (4- (1- methyl isophthalic acid H- pyrazoles -4- bases) pyridine -2- bases) -7- (2- picolines -4- Base) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

In room temperature to (4S)-N- (4- bromopyridine -2- bases) -9- methyl -7- (2- picoline -4- bases) -3,4- dihydro -1, 4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (300mg, 0.645mmol) are in 1,4- bis- Evil 1- methyl -4- (4,4,5,5- tetramethyl -1,3,2- dioxa boron is added in alkane (40mL) and the solution of the stirring in water (10mL) The amyl- 2- yls of heterocycle) -1H- pyrazoles (201mg, 0.967mmol) and K₃PO₄(411mg, 1.934mmol) and the 15min that deaerates.Then Add PdCl₂(dppf)-CH2Cl2 adducts (52.6mg, 0.064mmol) and the 5min that deaerates again, then stir 16h at 80 DEG C. (TLC eluant, eluents：Net ethyl acetate:Rf:0.3；UV activation).Reactant mixture is cooled to room temperature, frozen water is subsequently poured into simultaneously It is extracted with ethyl acetate (2x100ml).The organic layer of merging washs (50mL) with saturated brine solution, through anhydrous sodium sulfate drying And be evaporated in vacuo, obtain crude product.Crude compound (uses neutral alumina, in 60% acetic acid second through flash column chromatography Elute post in ester/petroleum ether), obtain pure compound (4S) -9- methyl-N- (4- (1- methyl isophthalic acid H- pyrazoles -4- bases) pyridine - 2- yls) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (126mg, 0.269mmol, 41.8% yield), it is white solid.LCMS(m/z):467.29[M+H]⁺；R_t =1.63min.

¹H NMR(400MHz,CDCl₃):δ 13.72 (s, 1H), 8.64-8.57 (m, 1H), 8.37 (dd, J=1.6, 0.8Hz, 1H), 8.30 (dd, J=5.2,0.8Hz, 1H), 8.21 (d, J=1.5Hz, 1H), 7.91 (d, J=0.8Hz, 1H), 7.85-7.80 (m, 1H), 7.76-7.70 (m, 1H), 7.41-7.36 (m, 1H), 7.10 (dd, J=5.2,1.6Hz, 1H), 5.70 (dd, J=6.0,3.1Hz, 1H), 3.96 (s, 3H), 3.22-3.08 (m, 3H), 3.03 (dd, J=12.0,3.2Hz, 1H), 2.75 (s, 3H), 2.51 (s, 3H), 2.34 (dq, J=13.3,6.5Hz, 1H), 2.08 (dt, J=14.4,7.9Hz, 1H).

Embodiment 371

Synthesize (4S) -8- chloro- N- (2- ((R) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (1- (2,2,2- trifluoros Ethyl) -1H- pyrazoles -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first Acid amides

At 0 DEG C to the chloro- N- of (4S) -8- (2- (((S) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridin-4-yl) -7- (1- (2,2,2- trifluoroethyls) -1H- pyrazoles -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diazaAdd in the solution of stirring of -5 (the 2H)-formamides (270mg, 0.455mmol) in methanol (5mL) Enter HCl/water solution (0.5ml, 6.00mmol), 1h is then stirred at room temperature.(TLC systems:10%MeOH/DCM, R_fValue：0.3). By reactant mixture saturation NaHCO₃It is water-soluble basified and extracted (2x50mL) with 10%MeOH/DCM.The organic layer of merging is used Sodium sulphate is dried and concentrated.Crude compound is through chromatogram purification (Grace instruments, C-18 reversed-phase columns, mobile phase A:0.1% formic acid/ Water；B：Acetonitrile, product is eluted in 60-65%B/A), obtain the chloro- N- of (4S) -8- (2- ((R) -2,3- dihydroxy propoxyl group) pyrroles Pyridine -4- bases) -7- (1- (2,2,2- trifluoroethyls) -1H- pyrazoles -4- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza- 5 (2H)-formamides (220mg, 0.393mmol, 86% yield), it is white solid.LCMS(m/ z):554.23[M+H]⁺, Rt=1.84min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.67 (s, 1H), 8.14 (d, J=9.21Hz, 2H), 7.96 (d, J= 5.70Hz, 1H), 7.62 (s, 1H), 7.13 (d, J=1.75Hz, 1H), 6.95 (dd, J=5.81,1.86Hz, 1H), 5.63 (dd, J=5.81,2.96Hz, 1H), 4.82 (q, J=8.33Hz, 2H), 4.47-4.43- (m, 2H), 4.27 (br d, J= 5.48Hz,1H),3.95-4.01(m,1H),3.62-3.69(m,2H),3.08-3.32(m,3H),3.03-2.96-(m,2H), 2.38-2.26-(m,1H),2.09-2.00-(m,1H)。

Embodiment 372

Synthesize (4S) -8- chloro- N- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (1- (2,2,2- trifluoros Ethyl) -1H- pyrazoles -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-first Acid amides

At 0 DEG C to the chloro- N- of (4S) -8- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridin-4-yl) -7- (1- (2,2,2- trifluoroethyls) -1H- pyrazoles -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diazaAdd in the solution of stirring of -5 (the 2H)-formamides (250mg, 0.421mmol) in methanol (5mL) Enter HCl/water solution (0.5ml, 6.00mmol), 1h is then stirred at room temperature.(TLC systems:10%MeOH/DCM, R_fValue：0.3). By reactant mixture saturation NaHCO₃It is basified and extracted (2x50mL) with 10%MeOH/DCM.The organic layer of merging is through sulphur Sour sodium is dried and concentrated.Crude compound is through chromatogram purification (Grace instruments, C-18 reversed-phase columns, mobile phase A:0.1% formic acid/ Water；B:Acetonitrile；Product is eluted in 59-64%B/A), obtain the chloro- N- of (4S) -8- (2- ((S) -2,3- dihydroxy propoxyl group) pyrroles Pyridine -4- bases) -7- (1- (2,2,2- trifluoroethyls) -1H- pyrazoles -4- bases) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3- B] [1,4] diaza- 5 (2H)-formamides (210mg, 0.374mmol, 89% yield), it is white solid.LCMS(m/ z):554.23[M+H]⁺, Rt=1.84min.

¹H NMR(400MHz,CDCl₃):δ ppm 12.67 (s, 1H), 8.14 (d, J=10.30Hz, 2H), 7.96 (d, J= 5.92Hz, 1H), 7.62 (s, 1H), 7.12 (d, J=1.75Hz, 1H), 6.95 (dd, J=5.81,1.86Hz, 1H), 5.63 (dd, J=5.92,3.07Hz, 1H), 4.82 (q, J=8.33Hz, 2H), 4.40-4.51 (m, 2H), 4.45 (dd, J=4.82, 1.75Hz, 1H), 3.99 (t, J=4.71Hz, 1H), 3.59-3.75 (m, 2H), 3.15-3.32 (m, 2H), 3.07-3.14 (m, 1H), 2.86-3.03 (m, 2H), 2.25-2.39 (m, 1H), 2.05 (dt, J=14.52,7.54Hz, 1H).

Embodiment 373

At 0 DEG C to the chloro- N- of (4S) -8- (2- (((R) -2,2- dimethyl -1,3- dioxolane -4- bases) methoxyl group) Pyridin-4-yl) -7- (6- (trifluoromethyl) pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] DiazaHCl/water is added in the solution of stirring of -5 (the 2H)-formamides (60mg, 0.102mmol) in methanol (10mL) molten Liquid (1mL, 32.9mmol), is then stirred at room temperature 2h.(TLC systems:10%MeOH/DCM, Rf value：0.25).By reaction mixing Thing saturation NaHCO₃It is water-soluble basified and be extracted with ethyl acetate (3x30mL).The organic layer of merging is dry through anhydrous sodium sulfate It is dry, filter and concentrate.Crude compound purifies (Grace instruments, anti-phase C-18 posts, with the formic acid of 27% acetonitrile/1% through column chromatography The aqueous solution is eluted), obtain the chloro- N- of (4S) -8- (2- ((S) -2,3- dihydroxy propoxyl group) pyridin-4-yl) -7- (6- (fluoroforms Base) pyridine -2- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (32mg, 0.058mmol, 57.0% yield), it is pale solid.LCMS(m/z):551.20[M+H]⁺, Rt= 1.99min。

¹H NMR(400MHz,CDCl₃):δ ppm 12.74 (s, 1H), 8.08 (t, J=7.78Hz, 1H), 7.98 (d, J= 7.89Hz, 1H), 7.91-7.80 (m, 2H), 7.69 (s, 1H), 7.24-7.03 (m, 1H), 6.81 (dd, J=5.92,1.97Hz, 1H), 5.65 (dd, J=5.81,3.18Hz, 1H), 4.43 (d, J=4.82Hz, 2H), 4.29 (d, J=5.92Hz, 1H), 3.96 (br d, J=5.26Hz, 1H), 3.70-3.46 (m, 2H), 3.25 (br dd, J=9.65,6.80Hz, 2H), 3.08-3.18 (m, 1H), 3.07-2.94 (m, 1H), 2.86 (dd, J=7.02,6.14Hz, 1H), 2.29-2.39 (m, 1H), 2.07 (dt, J= 14.47,7.02Hz,1H)

Embodiment 374

(4S)-N- (4- (1- ethyl -1H- pyrazoles -4- bases) pyridine -2- bases) -7- (2- picoline -4- bases) -3 is synthesized, 4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

Under argon gas to (4S)-N- (4- bromopyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- bridges Methylene pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (500mg, 1.108mmol) are in 1,4- dioxanes In (50mL) and water (10mL) K is added in the solution of degassing₃PO₄(705mg, 3.32mmol), 1- ethyls -4- (4,4,5,5- tetramethyls Base -1,3,2- dioxaborolan -2- bases) -1H- pyrazoles (320mg, 1.440mmol), 10min is then stirred at room temperature.So After add PdCl₂(dppf)-CH₂Cl₂Adduct (90mg, 0.111mmol) and 80 DEG C stir 3h.(TLC systems:10% first Alcohol/DCM, Rf values：0.5).Water (100mL) adds to reactant mixture and is extracted with ethyl acetate (3x100mL).By the extraction of merging Thing aqueous salt solu-tion (100mL) is taken, through anhydrous Na₂SO₄Dry, filter and filtrate evaporation is obtained into crude compound.Thick production Thing through chromatogram purification (neutral alumina, in DCM elute), obtain (4S)-N- (4- (1- ethyl -1H- pyrazoles -4- bases) pyridine - 2- yls) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (300mg, 0.641mmol, 57.9% yield), it is pale solid.LCMS(m/z):467.25[M+H ]⁺, Rt=1.57min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.56 (s, 1H), 8.62 (d, J=5.26Hz, 1H), 8.38 (dd, J= 1.43,0.77Hz, 1H), 8.31 (dd, J=5.26,0.66Hz, 1H), 8.26-8.15 (m, 1H), 7.92 (d, J=0.66Hz, 1H), 7.86 (s, 1H), 7.72 (dd, J=5.26,1.32Hz, 1H), 7.63 (d, J=8.11Hz, 1H), 7.49 (d, J= 8.11Hz, 1H), 7.25-7.03 (m, 1H), 5.71 (dd, J=5.81,3.18Hz, 1H), 4.23 (q, J=7.38Hz, 2H), 3.35-3.16 (m, 3H), 3.03 (dd, J=12.28,3.29Hz, 1H), 2.75 (s, 3H), 2.44-2.24 (m, 1H), 2.11 (dt, J=14.31,7.43Hz, 1H), 1.59-1.53 (m, 3H).

Embodiment 375

Synthesis (4S)-N- (4- (1H- pyrazoles -4- bases) pyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydros - 1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4S)-N- (4- bromopyridine -2- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (650mg, 1.440mmol), 4- (4,4,5,5- tetramethyl -1,3, 2- dioxaborolan -2- bases) -1H- pyrazoles (419mg, 2.160mmol) and Na₂CO₃(458mg, 4.32mmol) is in 1,4- Pd (Ph are added in dioxane (24mL) and water (6mL) in the solution of degassing₃P)₄(166mg, 0.144mmol), and in 80 DEG C of stirrings 16h.(TLC eluant, eluents：10%MeOH/ ethyl acetate, R_f- 0.4, UV activation).1,4- dioxanes are evaporated under reduced pressure, by acquisition Residue diluted with water (50ml) is simultaneously extracted with ethyl acetate (2x100ml).The organic layer aqueous salt solu-tion of merging (30mL), through anhydrous sodium sulfate drying and is evaporated under reduced pressure, and obtains crude product.Crude product is through flash column chromatography (using neutral Aluminum oxide, in 5% methanol/ethyl acetate elute), obtain desired product (4S)-N- (4- (1H- pyrazoles -4- bases) pyridine - 2- yls) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5 (2H)-formamide (150mg, 0.333mmol, 23.09% yield), it is pale solid.LCMS(m/z):439.21[M+H ]⁺,R_t=1.33min.

¹H NMR(400MHz,DMSO-d6):δ ppm 13.50 (s, 1H), 13.20 (br s, 1H), 8.59 (d, J= 5.26Hz, 1H), 8.42-8.31 (m, 3H), 8.25 (s, 1H), 8.06-7.93 (m, 2H), 7.81 (d, J=8.11Hz, 1H), 7.75-7.66 (m, 1H), 7.37 (dd, J=5.15,1.43Hz, 1H), 5.54 (dd, J=5.59,2.96Hz, 1H), 3.25- 3.09 (m, 3H), 2.98 (dd, J=11.95,3.18Hz, 1H), 2.64 (s, 3H), 2.35-2.18 (m, 1H), 2.06-1.89 (m,1H)。

Embodiment 376

Synthesize (4S)-N- (5- (2- Jia Ji oxazole -5- bases) pyridazine -3- bases) -7- (2- picoline -4- bases) -3,4- two Hydrogen -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4S)-N- (5- chlorine pyridazine -3- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (400mg, 0.981mmol) are in 1,4- dioxanes (16mL) and water 2- methyl -5- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- bases) is added in the solution of stirring in (4mL) Oxazole (308mg, 1.471mmol) and K₃PO₄(625mg,2.94mmol).By gained reactant mixture argon-degassed 15min. Pd (Ph are added thereto₃P)₄(113mg, 0.098mmol) and the 5min that deaerates again.By gained reactant mixture in 100 DEG C of stirrings 18h.(TLC eluant, eluents：10%MeOH/DCM, R_f- 0.6, UV activation).Reactant mixture is diluted with water and extracted with ethyl acetate Take (3 × 30mL).The organic layer of merging is with aqueous salt solu-tion (20mL), through anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure, Obtain crude compound.Crude compound connects through flash column chromatography (100-200 silica gel is eluted in 4%MeOH/DCM) And purify (Prep-HPLC conditions through preparation HPLC:MP-A:10mm ammonium hydrogen carbonate (aqueous solution) MP-B：Acetonitrile post： KinetexC8 (150*30) mm*5u methods：% ' B ':0/10,9/50,9.5/100,13/100,13.5/50,15/50 flow velocity： 30ml/min eluant, eluents：ACN+THF+MeOH temperature:Environment temperature), obtain desired compound (4S)-N- (5- (2- Jia Ji Evil Azoles -5- bases) pyridazine -3- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1, 4] diaza- 5 (2H)-formamides (124mg, 0.271mmol, 27.6% yield), it is pale solid.LCMS(m/z): 455.19[M+H]⁺,R_t=1.38min.

¹H NMR(400MHz,CDCl₃):δ ppm 13.65 (s, 1H), 9.01 (d, J=2.63Hz, 1H), 8.74 (d, J= 5.26Hz, 1H), 8.28 (d, J=2.63Hz, 1H), 7.79 (s, 1H), 7.70 (d, J=7.89Hz, 1H), 7.57 (s, 1H), 7.51 (d, J=5.26Hz, 1H), 7.42 (d, J=7.89Hz, 1H), 5.68 (dd, J=5.92,3.29Hz, 1H), 3.33- 3.16 (m, 3H), 3.06 (dd, J=12.28,3.29Hz, 1H), 2.72 (s, 3H), 2.60 (s, 3H), 2.45-2.29 (m, 1H), 2.17-2.03(m,1H)

Embodiment 377

Synthesize (4S) -7- (2- picoline -4- bases)-N- (5- (oxazole -5- bases) pyridazine -3- bases) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides

To (4S)-N- (5- chlorine pyridazine -3- bases) -7- (2- picoline -4- bases) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides (500mg, 1.226mmol) are in 1,4- dioxanes (16mL) and water 5- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- base) oxazoles are added in the solution of stirring in (4mL) (359mg, 1.839mmol) and K₃PO₄(781mg,3.68mmol).By gained reactant mixture argon-degassed 15min.Xiang Qi Middle addition Pd (Ph₃P)₄(142mg, 0.123mmol) and the 5min that deaerates again.Gained reactant mixture is stirred into 18h at 100 DEG C. (TLC eluant, eluents：10%MeOH/DCM, R_f- 0.6, UV activation).Reactant mixture is diluted with water and is extracted with ethyl acetate (3 ×30mL).The organic layer of merging is with aqueous salt solu-tion (20mL), through anhydrous Na₂SO₄Dry, filter and be concentrated under reduced pressure, obtain Crude compound.Crude compound is then passed through through flash column chromatography (100-200 silica gel is eluted in 4%MeOH/DCM) Preparation HPLC purifies (Prep-HPLC conditions:MP-A:10mm ammonium hydrogen carbonate (aqueous solution) MP-B：Acetonitrile post：Kinetex C8 (150*30) mm*5u methods：% ' B ':0/10,9/50,9.5/100,13/100,13.5/50,15/50 flow velocity：30ml/min is washed De- agent：ACN+THF+MeOH temperature:Environment temperature), obtain desired product (4S) -7- (2- picoline -4- bases)-N- (5- (oxazole -5- bases) pyridazine -3- bases) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza-5(2H)- Formamide (49mg, 0.111mmol, 9.02% yield), it is pale solid.LCMS(m/z):441.23[M+H]⁺,R_t= 1.32min。

¹H NMR(400MHz,CDCl₃):δ ppm 13.68 (s, 1H), 9.07 (d, J=2.41Hz, 1H), 8.75 (d, J= 5.04Hz, 1H), 8.35 (d, J=2.41Hz, 1H), 8.05 (s, 1H), 7.96 (s, 1H), 7.70 (d, J=7.89Hz, 1H), 7.57 (s, 1H), 7.51 (d, J=5.04Hz, 1H), 7.42 (d, J=7.89Hz, 1H), 5.69 (dd, J=6.03,2.96Hz, 1H), 3.39-3.13 (m, 3H), 3.07 (dd, J=12.28,3.07Hz, 1H), 2.72 (s, 3H), 2.50-2.29 (m, 1H), 2.11 (dt, J=14.25,7.13Hz, 1H).

At 0 DEG C to (4S)-N- ((R) -2,3- dihydroxypropyls) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydros -1,4- Endo-methylene group pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (135mg, 0.320mmol) are at ether (10mL) In stirring suspension in add HCl diethyl ether solution (2M) (0.160mL, 0.320mmol), 1h is then stirred at room temperature. (TLC eluant, eluents：10% methanol/DCM:R_f-0.1；UV activation).Then reactant mixture is concentrated under reduced pressure in nitrogen atmosphere, Obtain solid chemical compound.Solid chemical compound is dissolved in 10ml water and acetonitrile (1:1) in, it is concentrated under reduced pressure, obtains pure compound (4S)-N- ((R) -2,3- dihydroxypropyls) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene group pyridos [2,3-b] [1,4] diaza- 5 (2H)-carboxamide hydrochlorides (120mg, 0.256mmol, 80% yield), it is canescence Solid.LCMS(m/z):423.04[M+H]⁺,R_t=1.76min.

¹H NMR(400MHz,DMSO-d₆):δ 10.12 (t, J=5.04Hz, 1H), 8.38 (d, J=7.67Hz, 1H), 8.21 (s, 1H), 7.97 (d, J=7.89Hz, 1H), 7.89-7.71 (m, 3H), 5.59 (m, 1H), 3.68-3.27 (m, 8H), 3.26-3.15(m,1H),2.47-2.30(m,1H),2.19-2.02(m,1H)。

Embodiment 378

Synthesize (4S)-N- (2- hydroxyethyls) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene group pyrroles Pyridine simultaneously [2,3-b] [1,4] diaza- 5 (2H)-formamides

At 0 DEG C to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene group pyridos [2,3- B] [1,4] diazaIn the solution of the stirring of (0.3g, 0.983mmol) in THF (10mL) add triethylamine (0.822mL, 5.90mmol) and triphosgene (0.292g, 0.983mmol), 1h is then stirred at room temperature, add 2- ethylaminoethanols (0.172mL, 1.965mmol) and by reactant mixture at 80 DEG C stir 16h.(TLC eluant, eluents：Net ethyl acetate:R_f-0.5；UV activation). Make reactant mixture room temperature and be diluted with water, be extracted with ethyl acetate (2x30mL).The organic layer aqueous salt solu-tion of merging (2x20mL), through anhydrous Na₂SO₄It is dried, filtered and concentrated, obtains crude compound.Crude compound is purified through column chromatography (100-200 silica gel is eluted with 5% methanol/DCM), obtains desired compound (4S)-N- (2- hydroxyethyls) -7- (3- (three Methyl fluoride) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (186mg, 0.467mmol, 47.5% yield), it is light green color jelly liquid.LCMS(m/z):393.08[M+H]⁺,R_t= 1.89min。

¹H NMR(400MHz,DMSO-d₆):δ ppm 10.34 (d, J=10.52Hz, 1H), 8.31 (s, 1H), 8.17 (s, 1H),7.84-7.76(m,1H),7.76-7.59(m,3H),5.48-5.40(m,1H),3.63-3.47(m,2H),3.45-3.30 (m,2H),3.28-2.89(m,4H),2.30-2.08(m,1H),1.95-1.75(m,1H)。

Embodiment 379

Synthesize (4S) -7- (3- (trifluoromethyl) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyridos [2,3-b] [1,4] Diaza- 5 (2H)-formamides

Room temperature to (4S) -7- (3- (trifluoromethyl) phenyl) -2,3,4,5- tetrahydrochysene -1,4- endo-methylene groups pyrido [2, 3-b] [1,4] diazaIn the solution of (250mg, 0.819mmol) in THF (25mL) add triethylamine (0.685ml, 4.91mmol), triphosgene (243mg, 0.819mmol) and stirring 30min are subsequently added into.Then the THF solution of 2M ammonia is added (1.228mL, 2.457mmol) and reactant mixture is heated into 16h at 65 DEG C.(TLC eluant, eluents：70%EtOAc/ hexanes:R_f- 0.3；UV activation).Reactant mixture is cooled to room temperature, is concentrated in vacuo and by residue distribution in water (30mL) and EtOAc Between (2x35mL).Separate organic layer and through anhydrous sodium sulfate drying, filter and filtrate evaporation is obtained into crude compound.Thick production Thing is through flash column chromatography (neutral alumina, eluant, eluent：50% ethyl acetate/hexane), obtain (4S) -7- (3- (fluoroforms Base) phenyl) -3,4- dihydro -1,4- endo-methylene groups pyrido [2,3-b] [1,4] diaza- 5 (2H)-formamides (225mg, 0.644mmol, 79% yield), it is white solid.LCMS(m/z):349.08[M+H]⁺,R_t=1.97min.

¹H NMR(400MHz,CDCl₃):δ ppm 10.23-10.02 (m, 1H), 8.00 (s, 1H), 7.94 (d, J= 7.89Hz, 1H), 7.69-7.65 (m, 1H), 7.62-7.54 (m, 2H), 7.29 (d, J=7.89Hz, 1H), 5.63 (dd, J= 6.03,3.18Hz, 1H), 5.23 (br s, 1H), 3.31-3.08 (m, 3H), 2.96 (dd, J=11.84,3.29Hz, 1H), 2.28 (dddd, J=14.03,9.98,6.03,3.95Hz, 1H), 2.10-2.00 (m, 1H).

The total length SIRT1 of embodiment 380. generation

Expression total length people SIRT1 (hSIRT1) albumen is allowed to carry C- ends His₆Label and such as Hubbard. et al. (2013) Science 339, are purified described in 1216.Each cell paste is suspended in containing 1,000U again on ice Buffer A (50mM Tris-HCl pH 7.5, the 250mM NaCl, 25mM of Benzonase nucleases (Sigma Aldrich) Imidazoles and 0.1mM TCEP), it is supplemented with the complete protease inhibitors mixing tablet (Roche) without EDTA.Cell passages through which vital energy circulates Ultrasonication is rushed, is opened in 40W with 50% and 50% stops carrying out 12 minutes altogether.Insoluble fragment is removed by centrifugation. The supernatant of clarification is directly loaded to 1mL HisTrap FF Crude posts (GE Lifesciences).Washed with buffer A Afterwards, SIRT1 is washed with buffer B (50mM Tris-HCl pH 7.5,250mM NaCl, 500mM imidazoles and 0.1mM TCEP) It is de-.By the albumen further with size exclusion chromatography buffer solution C (50mM Tris-HCl pH 7.5,300mM NaCl, 0.1mM TCEP) in purified, its use the posts of Hi-load Superdex 200 16/60 (GE Lifesciences).Enzyme Concentration is determined with the Bradford analyses using BSA as standard items.Final purity of protein colloid light densitometry It is estimated.Albumen is confirmed by LC/MS.All albumen are all higher than 90% purity.

Embodiment 382.SIRT1 deacetylations

SIRT1 deacetylations be 25 DEG C of reaction buffers (50mM HEPES-NaOH, pH7.5,150mM NaCl, 1mM DTT and 1%DMSO) middle generation (Smith, the B.C. for carrying out, using continuation PNC1/GDH coupled assays monitoring niacinamide Et al. (2009) Anal Biochem 394,101) or with mass spectrum monitor O- acetyl group ADP ribose (OAcADPr) generation (Hubbard. et al. (2013) Science 339,1216).The ultimate density of used PNC1/GDH coupling systems composition is 20 units/mL oxen GDH (Sigma-Aldrich), 1uM yeast PNC1,3.4mM α-ketoglutaric acids and 220 μM of NADH or NADPH. Use 6.22mM^-1cm^-1Extinction coefficient and 0.81cm optical path lengths change absorbance at 340nm, obtain 150uL reactions used Production concentration.The analysis that monitoring OAcADPr is produced is carried out and by using termination in the reaction buffer containing 0.05%BSA Solution is quenched the deacetylation and takes time point, and the stop bath provides the ultimate density of 1% formic acid and 5mM niacinamide.Quench The reactant mixture gone out uses 1:1 acetonitrile:5 times of methanol dilution simultaneously rotates 10 minutes for protein precipitation with 5,000x g, afterwards will It is analyzed with the high flux mass spectrometer systems (Agilent, Wakefield, MA) of Agilent RapidFire 200, the mass spectrometer system It is coupled to the mass spectrographs of ABSciex API 4000 equipped with electric spray ion source.Respectively from American Century Peptide And Biopeptide, Inc. obtain Ac-p53 (W5) (Ac-RHKK based on p53^AcW-NH₂) and TAMRA (Ac-EE-K (biologies Element)-GQSTSSHSK (Ac) NleSTEG-K (5TMR)-EE-NH₂) peptide.Substrate K_MMeasure is by the of fixed saturated concentration In the case of two substrates, change what a concentration of substrate was carried out.SIRT1 is activated and inhibition analysis is at 25 DEG C, containing 0.05% Carry out and analyzed using OAcADPr in BSA reaction buffer.Before substrate is added, enzyme and compound are carried out 20 minutes precultures.For total length hSIRT1 activation screening, compound determines its dose response in duplicate.In order to K_M- The activating agent of regulation is sensitive, uses their K_MAbout 1/10th concentration of substrate of value.Determine the dose-dependant of 5 compounds The property and multiple-activation data equation 1 is described.

Wherein v_x/v₀It is the presence of (v_x) with (v is not present₀) activator (X) reaction rate ratio, RV_maxIt is infinite Relative speed under big activator concentration, EC₅₀It is the RV of generation 1/2nd_maxThe concentration and b of required activator are v_x/v₀ Minimum value.

The biochemical activity of embodiment 383.

Based on the mass spectrographic conditioning agent analyzed for identifying SIRT1 activity.The analysis based on TAMRA is used with as follows The peptide of 20 shown amino acid：

Ac-EE-K (biotin)-GQSTSSHSK (Ac) NleSTEG-K (5TMR)-EE-NH₂(SEQ ID NO:1),

Wherein K (Ac) is the lysine residue of acetylation, and Nle is nor-leucine.The peptide is in C- ends fluorogen 5TMR (exciting 540nm/ to launch 580nm) mark.The sequence of the peptide substrates is based on the p53 modified with several places.In addition, using nor-leucine Instead of the methionine residues being naturally occurring in sequence because methionine during synthesis and purifying to oxidation-sensitive.Base The peptide with amino acid as follows is used in Trp analysis：

Ac-R-H-K-K(Ac)-W-NH₂(SEQ ID NO:2),

Mass spectral analysis based on TAMRA is carried out as follows：At 25 DEG C, reaction buffer (50mM Tris- acetates, pH8, 137mM NaCl, 2.7mM KCl, 1mM MgCl₂, 5mM DTT, 0.05%BSA) in, by 0.5 μM of peptide substrates and 120 μM of β NAD⁺ It is incubated 25 minutes with 10nM SIRT1.SIRT1 albumen is obtained as follows：By SirT1 gene clonings to the carrier containing T7- promoters In, then it is converted and expressed in BL21 (DE3) bacterial cell.Test compound is added to the reaction with various concentration In mixture, obtained reaction is monitored.After being incubated 25 minutes with SIRT1,10 μ L 10% formic acid is added with terminating reaction.Will The reactant mixture sealing arrived, and freeze the mass spectral analysis after being used for.The NAD- mediated by Sirtuin is determined to rely on Property deacetylation formation deacetylated peptide substrate amount (or, the O- acetyl group-ADP- ribose (OAADPR) of generation Amount), can accurately measure in the presence of the test compound of various concentration relative to the control reaction for lacking test compound Relative SIRT1 activity.

Trp mass spectral analyses carry out as follows：At 25 DEG C, in reaction buffer (50mM HEPES pH 7.5,1500mM NaCl, 1mM DTT, 0.05%BSA) in, by 0.5 μM of peptide substrates and 120 μM of β NAD⁺It is incubated 25 minutes with 10nM SIRT1. SIRT1 albumen is obtained as follows：By SirT1 gene clonings into the carrier containing T7- promoters, then in BL21 (DE3) bacterium Express, and purify in cell, as described in further detail below.Test compound is added to the reactant mixture with various concentration In, monitor obtained reaction.After being incubated 25 minutes with SIRT1,10 μ L 10% formic acid is added with terminating reaction.It is anti-by what is obtained Answer mixture to seal, and freeze the mass spectral analysis after being used for.Then taken off by determining by NAD- dependence Sirtuins The amount (or, the amount of the deacetylated Trp peptides of generation) of the O- acetyl group-ADP- ribose (OAADPR) of acetylization reaction formation is come The relative SIRT1 of measurement is active (relative to the control reaction for lacking test compound in the presence of the test compound of various concentration). Test agent activates deacetylated degree expression for EC by SIRT1_1.5(that is, relative to the control for lacking test compound, Make the compound concentration required for SIRT1 activity increase by 50%), and maximum activation percentage (that is, obtains for test compound Relative to control (100%) maximum activity).

When reacting beginning, negative control is used as (for example, it is allowed to determine maximum heavy by adding 1 μ L 500mM niacinamide Silent regulatory protein suppresses) carry out the control of Sirtuin activity suppression.The control of the activation of Sirtuin activity It is carried out as follows：Using 10nM Sirtuins, compound is replaced with 1 μ L DMSO, in the range of linearity of analysis, Given point in time determines the deacetylated amount of substrate.The time point is identical with time point used in test compound, and In the range of linearity, the change of end points representation speed.

For above-mentioned analysis, SIRT1 protein expressions are simultaneously purified as follows.By SirT1 gene clonings to containing T7- promoters In carrier, and it is transformed into BL21 (DE3).At 18 DEG C, by using 1mM IPTG (being used as N- terminal His-tags fusion protein) Induce to express the albumen, overnight, and harvested under 30,000x g.In lysis buffer (50mM Tris-HCl, 2mM Tris [2- carboxyethyls] phosphine (TCEP), 10 μM of ZnCl₂, 200mM NaCl) in, cell is cracked with lysozyme, and further with ultrasound Processing 10 minutes, so as to cracking completely.With Ni-NTA posts (Amersham) purifying protein, and merge the fraction containing pure protein, Concentration, makes its operation pass through fractionation column (Sephadex S200 26/60global).The peak containing soluble protein is collected, and Operation passes through ion exchange column (MonoQ).Gradient elution (200mM-500mM NaCl) obtains pure protein.By the protein concentration, And carry out Dialyse overnight with dialysis buffer (20mM Tris-HCl, 2mM TCEP).By albumen decile, and it is preceding -80 using DEG C freezing.

Use above-mentioned analysis, the compound of the regulation Sirtuin of identification activation SIRT1 formula (I), such as table 1 below institute Show.EC_1.5Value represents the concentration for 150% test compound activated for causing SIRT1.The EC of the activating compounds of formula (I)_1.5Value By A (EC_1.5<1μM)、B(EC_1.5：1-25μM)、C(EC_1.5>25 μM) represent.Maximum activation folds percentage is by A (activation folds >=150%) or B (activation folds<150%) represent." NT " represents not test；" ND " represents not detecting.The chemical combination numbered in table Thing is since compound number 10, and the corresponding STAC numbering systems and embodiment 90-106 for being Fig. 4 of bracket numbering (#) is (i.e., Compound number 68 is also STAC 1, therefore is denoted as 68 (1), also STAC：546(3)、444(4)、314(5)、816 (7), 76 (8) and 81 (9)).

It is different by two kinds it is worth noting that, the compound of the present invention is named by two kinds of different chemical names rules Chemistry drawing and/or chemical name computer program are produced, i.e., by Chem Axon (JChem-Excel) and Cambridge SofRespective company produces.

Table 1 (ISOMER=isomers)

Embodiment 375

The present invention relates to Sirtuin conditioning agent, it becomes known for improving cell survival in scientific literature, and Treatment and/or a variety of diseases and obstacle of prevention wide scope, the disease and obstacle include but is not limited to, for example, with aging or The relevant disease of stress reaction or obstacle, diabetes, obesity, neurodegenerative disease, angiocardiopathy, blood coagulation disorder, Inflammation, cancer and/or flush, and benefit from mitochondrial active increased disease or obstacle

In addition to treatment potentiality, SIRT1 activity and the structure and biology that are activated by small molecule Sirtuin conditioning agent Physical study is additionally operable to promote biological function, the mechanism of action of Sirtuin activation and the auxiliary to Sirtuin The understanding of the determination method of the novel Sirtuin conditioning agent of exploitation identification.

Based on above, documents below bibliography is refer to respectively, and egg is adjusted to prove the compounds of this invention as silence The effect of white conditioning agent and its associated with various diseases, such as below with reference to as illustrated in document or openly：

1.Marcia C.Haigis and David A.Sinclair,Mammalian Sirtuins:Biological Insights and Disease Relevance,Annu Rev Pathol.2010；5:253–295.

Haigis and Sinclair teachings：

" aging with multiple organ system health function reduction, cause by type ii diabetes, neurodegenerative disease, cancer and Morbidity and mortality rise caused by the diseases such as angiocardiopathy.In history, researcher is absorbed in investigation isolated organ Individual approach, be used as determine disease root strategy, it is desirable to design more preferable medicine.The research of the aging in yeast is produced The number of ways of the referred to as family of the conservative enzyme of Sirtuin, its life-span for influenceing increase organism and holistic health.From Since first known mammal Sirtuin SIRT1 being found before 10 years, our enzymes to Sirtuin Scientific principle solution, its regulation and control and its extensive physiological mammal and the ability of healthy span of improving have major progress.This summary is summarized And discuss over 10 years discovery progress and the coming years by facing challenges (see " summary " therein and bibliography).

2.Gizem Donmez1 et al., SIRT1 and SIRT2:emerging targets in neurodegeneration,EMBO Mol Med(2013)5,344–352.

Gizem Donmez1 et al. are instructed：

" Sirtuin is known with confrontation age-related disease such as cancer, diabetes, cardiovascular and neurodegeneration Disease, the NAD dependence protein deacetylases with protective effect.In mammal, there are seven kinds of Sirtuins (SIRT1-7), its diversity for showing Subcellular Localization and function.Although SIRT1 due to life it is preliminary contact and Heat limitation participation and be widely studied, the important biomolecule of other Sirtuins and therapeutic action are just recognized recently Can.Herein, we have looked back the latent effect and influence of SIRT1 and SIRT2 in neurodegenerative disease.We discuss SIRT1 and SIRT2 is in various neurodegenerative diseases (including Alzheimer disease (AD), Parkinson's (PD) and Huntington disease (HD) difference in functionality and target in).We also reported effects of the SIRT1 in neuron differentiation (due to that may be involved in In neurodegenerative conditions), and the potential therapeutic value using SIRT1 and SIRT2 in these diseases prospect as conclusion (see it In " summary " and bibliography).

3.Bracke et al., Targeted silencing of DEFB4 in a bioengineered skin- humanized mouse model for psoriasis:development of siRNA SECosome-based novel therapies；Exp Dermatol.In March, 2014；23(3):199-201.doi:10.1111/exd.12321.

Specifically, Bracke et al. is instructed：

" psoriasis is a kind of complicated inflammatory dermatoses, with extensive clinical manifestation.Human β-defensin-2 (hBD-2) Highly raised in psoriatic lesions, and be defined as the biomarker of disease activity.We pass through the biology in psoriasis SECosomes of the topical application containing DEFB4-siRNA has inquired into targeting hBD-2's in the skin of engineering-peopleization mouse model Potential benefit.By histological examination it was observed that in the significant improvement of psoriatic phenotype, skin texture normalization and dermal compartment The reduction of blood vessel number and size.Treatment causes trans glutaminase active, the expression of silk polyprotein and cuticula outward appearance to return to The level similar to the level found in regular regeneration application on human skin.The acquisition of the reliable skin peopleization mouse model of psoriasis with And may provide valuable preclinical work for the therapy target of the potential this disease of identification using SECosome technologies Tool.”

4.Karline Guilloteau et al., Skin Inflammation Induced by the Synergistic Action of IL-17A,IL-22 Recapitulates Some Features of psoriasis Oncostatin M, IL-1a,and TNF-a,J Immunol 2010；184:5263-5270.

Guilloteau et al. is instructed:

" keratinocyte plays key effect in regulation scytitis in response to environment and immunocyte stimulation aspect. The soluble factor that they are produced can be worked or be directly acted on immunocyte in the way of autocrine or paracrine Invasion and attack person.The activity of 36 kinds of cell factor Human Keratinocytes gene expression is screened, IL-17A, IL-22, carcinostatin is identified M, TNF-α and IL-1a as induced skin inflammation potential cell factor.This five kinds of proinflammatory cytokine collaboration increase CXCL8 With the generation of b- alexins 2 (BD2).In addition, showing for the vitro study of human skin explant in response to cell factor Like combinations, BD2, S100A7 and CXCL8 up-regulated expression.This 5 kinds of cell factors of internal intracutaneous injection increase in mouse CXCL1, CXCL2, CXCL3, S100A9 and BD3 are expressed, relevant with neutrophil cell infiltration.We demonstrate that and extending this Cooperative effect, using quantitative real-time PCR analysis, it was observed that the expression increase of 9 kinds of chemotactic factor (CF)s and 12 kinds of antimicrobial peptides.At this The keratinocyte that plant stimulates in the presence of combination of cytokines produces CXCL, CXCL5 and CXCL8 and increased neutrophil(e) granule is thin Born of the same parents' chemotactic activity is related.Similarly, BD2, BD3 and S100A7 high yield are related to increased antimicrobial acivity.Finally, The transcriptional profile observed in the external model of inflammation keratinocyte is related to damaging psoriatic skin.Ours grinds Study carefully result and show IL-17A, IL-22, carcinostatin M, the important enhancing activity of TNF-α and IL-1a Human Keratinocytes.This Especially meaningful under the background of psoriasis, wherein these cell factors are over-expressed and can acted synergistically with chemotactic factor (CF) Played a significant role in the up-regulation produced with antimicrobial peptide.The Journal of Immunology, 2010,184：5263- 5270 (see " summary " therein and bibliography) ".

The measure description of embodiment 376

PBMC is determined

Sirtuin 1 (Sirt1) is silent message conditioning agent 2 (Sir2) homologue, is NAD dependence Group IIIs The member of histone deacetylase.Sirt1 to histone, transcription factor and it is nonhistones on lysine residue carry out it is deacetylated Change.Sirt1 has been demonstrated to participate in aging, Cycle Regulation, Apoptosis, Metabolism regulation and inflammation.Sirt1 activation causes The lysine 310 of the RelA/p65 subunits of nuclear factor kappaB (NF-kB) transcription factor is deacetylated, its suppress NF-kB transcription and under Adjust the level of TNF α.TNF α is a kind of pleiotrophic cytokine, is mainly produced by macrophage and monocyte.TNF α is participated in closely Immune defense and chronic inflammation include psoriasis.The expression increase of 1 cytokines such as TNF α in known psoriatic skin, and Played an important role in the teiology of psoriasis (Uyemura K et al., 1993, J.Invest Dermatol, 101, p701). Importantly, anti-TNF agent is clinically used for psoriasis.Therefore, the Sirt1 of TNF α expression reduction is activated in induction inflammatory cell Agent should have therapeutic action in into severe psoriatic.

The measure based on PBMC/TNF α cells is developed, to identify inhibition response in PMBC (PBMC) Lipopolysaccharides (LPS) stimulates and discharged the Sirt1 of TNF α activator.In brief, PBMC is stimulated by LPS, causes TNF α to be secreted The increase of generation.TNF α protein level is measured by TNF α HTRF (homogeneous phase time discrimination fluorescence) kit (CisBio, Inc). Cell cracking and TNF α detection are carried out according to the explanation of manufacturer.Sirt1 activator is tested in the presence of LPS right to assess its The inhibitory action of TNF α release, and determine IC50 in dosage-response experiment.

Beta-alexin 2 (bD2) is determined

Sirtuin is the family of NAD dependence deacetylases, and it has extensive physiological function, and has related to And many autoimmunities and metabolic disease, including rheumatoid arthritis and type i diabetes.SIRT1 substrate is varied , and it is included in the inflammatory components that there is determination to act in natural and adaptive immune response, such as NF κ B, AP-1, FOXO And p53.

Psoriasis is the chronic inflammatory skin disease caused by autoimmunity and environmental factor by heredity.Lesion is characterised by The infiltration of the hyper-proliferative and inflammatory cell of keratinocyte in epidermis, causes the chronic erythema patch of white squama covering.In the past Research show that, by directly suppressing STAT3 acetylations, SIRT1 can prevent IL-22 (a kind of crucial in psoriasis Cell factor) effect (Sestito et al., 2011).In addition, SIRT1 is overexpressed and resveratrol treats (SIRT1 activation) all Keratinocyte differentiation (Blander et al., 2009) can be induced.

Beta-alexin 2 (bD2) is a kind of antimicrobial peptide, can be used as memory t cell, prematurity from epithelial secretion BMDC and the chemoattractant of neutrophil cell.Therefore, bD2 is the major part of scytitis reaction.With it is normal Skin is compared, and bD2 is not only induced in the damage epidermal cell of psoriatic, and bD2 is also that psoriatic's disease is tight Serum biomarkers (Jansen et al., 2009 of weight degree；Kamsteeg etc., 2009).In addition, bD2 may be with psoriasis Genetic correlation, because a nearest research discloses increased β-resist between plain plain gene copy number and psoriasis risk Significantly association (Hollox et al., 2008).It is worth noting that, bD2siRNA local delivery causes psoriasis Bioengineered Skin-peopleization mouse model in regular skin structure and protein expression recovery (Bracke et al., 2014).

Be previously described produce with simulate psoriasis inflammation external keratinocyte inflammation determine (Guilloteau et al., 2010；Teng etc., 2014).In these researchs, the cell factor mixing of IL-1 α, IL-17A, IL-22, OSM and TNF α is found Thing (being referred to as " M5 ") acts synergistically to produce " psoriasis inflammation shape " transcription spectrum in vitro in primary Human keratinocytes.At this In a little researchs, bD2 is to induce one of keratinocyte inflammatory reaction most strong respondent.

Therefore, this measure is further developed, is played a game the treatment of portion's psoriasis program with assessing SIRT1 activation immunomodulator compounds Effect.Specifically, for having carried out bar with the immortal human keratinocyte cell line (HaCaT) of M5 combination of cytokines extracorporeal treatments Piece optimization, to induce psoriasis inflammation (as described in bibliography above).Within the time of 48 hours, pass through bD2 ELISA The bD2 secretions for determining (Alpha Diagnostics) measurement are dramatically increased compared with the keratinocyte not stimulated.It can lead to Known compound (or it is important that with a small group SIRT1 activator) processing for suppressing psoriasis inflammation for the treatment of is crossed to suppress this BD2 is induced.Meanwhile, the cytotoxicity in 48 hours measure is true by CellTiter-Glo luminescent cells vitality test (Promega) It is fixed, to determine whether toxicity may work in bD2 responses.

Bibliography：

Blander G,Bhimavarapu A,Mammone T,Maes D,Elliston K,Reich C,Matsui MS,Guarente L,Loureiro JJ.SIRT1promotes differentiation of normal human keratinocytes.J Invest Dermatol.2009Jan；129(1):41-9.

Bracke S,Carretero M,Guerrero-Aspizua S,Desmet E,Illera N,Navarro M, Lambert J,Del Rio M.Targeted silencing of DEFB4in a bioengineered skin- humanized mouse model for psoriasis:development of siRNA SECosome-based novel Therapies.Exp Dermatol.2014 March；23(3):199-201.

Guilloteau K,Paris I,Pedretti N,Boniface K,Juchaux F,Huguier V, Guillet G,Bernard FX,Lecron JC,Morel F.Skin Inflammation Induced by the Synergistic Action of IL-17A,IL-22,Oncostatin M,IL-1alpha,and TNFalpha Recapitulates Some Features of Psoriasis.J Immunol.2014 March.

Jansen PA,Rodijk-Olthuis D,Hollox EJ,Kamsteeg M,Tjabringa GS,de Jongh GJ,van Vlijmen-Willems IM,Bergboer JG,van Rossum MM,de Jong EM,den Heijer M, Evers AW,Bergers M,Armour JA,Zeeuwen PL,Schalkwijk J.Beta-defensin-2protein is a serum biomarker for disease activity in psoriasis and reaches biologically relevant concentrations in lesional skin.PLoS One.2009；4(3): e4725。

Kamsteeg M,Jansen PA,van Vlijmen-Willems IM,van Erp PE,Rodijk-Olthuis D,van der Valk PG,Feuth T,Zeeuwen PL,Schalkwijk J.Molecular diagnostics of psoriasis,atopic dermatitis,allergic contact dermatitis and irritant contact dermatitis.Br J Dermatol.2010Mar；162(3):568-78.

Sestito R,Madonna S,Scarponi C,Cianfarani F,Failla CM,Cavani A, Girolomoni G,Albanesi C.STAT3-dependent effects of IL-22in human keratinocytes are counterregulated by sirtuin 1through a direct inhibition of STAT3 acetylation.FASEB J.2011Mar；25(3):916-27.

Teng X,Hu Z,Wei X,Wang Z,Guan T,Liu N,Liu X,Ye N,Deng G,Luo C,Huang N,Sun C,Xu M,Zhou X,Deng H,Edwards CK 3rd,Chen X,Wang X,Cui K,Wei Y,Li J.IL- 37ameliorates the inflammatory process in psoriasis by suppressing Proinflammatory cytokine production.J Immunol.2014 2 months 15；192(4):1815-23.

Yin Xiebing ＆IL-17

Psoriasis be with the chronic of multifactor pathogenesis, recurrent, Inflammatory Autoimmune skin disease, by heredity, Environment and immunopathogenesis factor influence (Griffiths CE et al., Lancet 2007；370：263-71).The feature of psoriasis It is recurrent outbreaks, raised, boundary clear the avette patch of erythema of the silvery scales with attachment.In histology, silver bits Disease is characterized in the presence of the nucleation keratinocyte layer thickened, is dashed forward (rete pegs) with the nail exaggerated, this is by cutin shape T cell, neutrophil cell and the skin infiltration of BMDC of hyper-proliferative and activation into cell cause (Schon MP N.Engl.J.Med.352：1899-1912).

The evidence of accumulation shows, the disease that psoriasis is mediated as Th17, by its characteristic cytokines IL-17A, IL- 17F and IL-22 drivings.IL-22 induce keratinocyte propagation, and IL-17A stimulate keratinocyte secrete chemotactic because Son and other pro-inflammatory mediators, other described pro-inflammatory mediators raise other inflammatory cells, including neutrophil cell, BMDC With congenital lymphoid cell (Martin DA et al., J Invest Dermatol 2013；133：17-26).

Clinical verification of the IL-17 approach in mediation psoriasis is to study to confirm by successful Ph3, and the research is shown Use significantly improving (Langley et al., NEJM 2014) for the mab treatment disease for targetting IL-17.In addition, in IL- 17 suppress after psoriatic lesions in overall transcryption analysis of spectrum will be a variety of from keratinocyte and leucocyte subgroup Inflammatory factor suppresses to similar level (Russell et al., the J Immunol 2014,192 with being observed in non-damaging skin： 3828-3836).In a word, these find to support effects of the IL-17 in mediation psoriatic pathogenesis.

Method (isolated skin measure)

The immunocyte being resident using Th17 cytokine mixtures in excised human skin's explant moderate stimulation skin, is led The notable up-regulation of Th17 relevant cell factors (IL-17A, IL-17F and IL-22) is caused, the system is established as organizing based on people by it Psoriasis model.After Th17 cytokine mixtures are stimulated, test compound regulation IL-17A, IL-17F and IL-22 are assessed The ability of expression, uses isolated skin cultural method.

Briefly, the excised human skin obtained that performed the operation from abdominoplasty is processed, to remove fat, organizes quilt Subcutaneously cut into about 750 microns.Then the skin of cutting is connected twice in the room temperature PBS containing antibiotic/antimycotic solution Continuous to rinse cleaning 5-10 minutes, the circle that skin biopsy cuts into 10mm diameters with disposable special purpose biopsy punch is cut In piece, the upper chambers for then placing it in 0.4 μm of PCF films Transwell (Millicell#PIHP01250), it contains 30 μ The 64% bovine collagen solution (Organogenesis, #200-055) that l is prepared with keratinization culture medium.By skin samples at 37 DEG C Under in humidifying chamber on collagen solution place 30 minutes.Skin samples on Transwell are transferred to 6 orifice plates (per 1, hole Sample) in, and bottom compartment is full of 1ml complete mediums (keratinization culture medium).

First day after belly plastic operation, skin explant is cultivated in keratinization culture medium, and is incubated at 37 DEG C Overnight.Specifically, with Th17 mixtures (CD3,1 μ g/ml, CD28,2 μ g/ml, IL-1b, 10ng/ml, IL-6,5ng/ml, TGFb, 1ng/ml, IL-21,10ng/ml, anti-IL-4,1 μ g/ml and anti-INFg, 1 μ g/ml) stimulate application on human skin explant (N=3/ Condition).Add 1,3 and 10uM test compound simultaneously with Th17 mixtures.Collect tissue within 24 hours after Th17 activation, point It is used to carry out quantitative (IL-17A, IL-17F, IL-22) transcript using qPCR from RNA.

Using Qiagen Mini RNA separating kits (catalog number (Cat.No.) 74106) total serum IgE is separated from about 40mg tissue. In short, in Precellys-24 machines, the RLT buffer solutions of 1%2- β-mercapto-ethanol are supplemented with using 300 μ l, Tissue is shredded and homogenized 30 seconds under 6300rpm, 10 cycles are run, wherein there is the cooling suspending period (ice of 2 minutes break).The water that 490 μ l contain 10 μ l Proteinase Ks is added into homogenate, and is digested 15 minutes at 55 DEG C.By the tissue of digestion In 10,000G centrifugal sedimentations 3 minutes with sedimentation cell fragment, and using Qiagen RNeasy micro-columns according to the side of manufacturer Case separates supernatant for RNA.Using the quantitative total serum IgEs of Nanodrop 2000, and (enclosed in Agilent biological analysers File) on analyzed.Using Invitrogen SuperScript VILO cDNA synthetic agent box (#11754-050), 1.4 μ gRNA are used in 20 μ lPCR volumes as template to produce cDNA templates.Then for subsequent qPCR by cDNA with 1: 25 dilutions, treat that quantitative gene uses specific TaqMan probe for each, are quantified.Use Delta Delta CT formula Calculate the relative expression of the rna level of gene interested.

Equivalent

The invention provides compound, pharmaceutical composition, method, purposes of regulation Sirtuin etc..Although The specific embodiment of theme invention is discussed, but description above is illustrative, is not restricted.Reading this explanation After book, many changes of the invention are obvious for a person skilled in the art.The four corner of the present invention should be by right The gamut of claim and its equivalent, and specification and the change are determined.

Claims

1. formula (I) compound or its pharmaceutically acceptable salt, formula (I) compound is:

Wherein：

X₁Or X₂It is independently selected from-N or-C；

R¹Heterocyclic radical, aryl, heteroaryl ,-C (O) R replaced for hydrogen, halogen ,-CN, carbocylic radical, heterocyclic radical ,-N-_aOr-C (O)- NR_bR_c；

R⁴For hydrogen or-C (O) NR_bR_c；

Wherein：

Work as X₂When being-N, R₂It is not present；Or

Work as X₂When being-C, R₂As defined above；

Each R as defined above¹、R²、R³And R⁴Optionally further it is selected from following one or more substituents substitution：Hydrogen, halogen Element ,-OH ,-(CH₂)_xOH、-C≡N、-NR_dR_e, straight or branched C₁-C₆Alkyl, straight or branched-C₁-C₆Haloalkyl, straight chain Or side chain C₁-C₆Alkoxy, straight or branched C₁-C₆Halogenated alkoxy ,-O- straight or brancheds-C₁-C₆Haloalkyl ,-C₁-C₆Ring Alkyl ,-(CH₂)_x- cycloalkyl, heterocyclic radical, aryl ,-heteroaryl ,-(CH₂)_x- heteroaryl ,-O- (CH₂)_xCH(OH)CH₂(OH), Or-C (O) OR_f；

Each R as defined above_a、R_b、R_c、R_d、R_eOr R_fIt is independently selected from hydrogen, straight or branched C₁-C₆Alkyl, straight or branched-C₁-C₆ Haloalkyl ,-C₁-C₆- cycloalkyl ,-(CH₂)_xC₁-C₆- cycloalkyl, heterocyclic radical ,-N- heterocyclic radicals, aryl, heteroaryl ,-(CH₂)_x Heteroaryl or-(CHR_g)_xHeteroaryl；

Wherein：

Each R as defined above_a、R_b、R_c、R_d、R_eOr R_fOptionally further it is selected from following one or more substituents substitution： Hydrogen, halogen ,-OH ,-C ≡ N, straight or branched C₁-C₆Alkyl, straight or branched-C₁-C₆Haloalkyl, straight or branched C₁-C₆ Alkoxy ,-O- straight or brancheds-C₁-C₆Haloalkyl ,-C₁-C₆Cycloalkyl, carbocylic radical ,-(CH₂)_x- carbocylic radical ,-heterocyclic radical ,- O- heterocyclylalkyls ,-heteroaryl ,-(CH₂)_x- heteroaryl ,-O- (CH₂)_xCH(OH)CH₂(OH)、-(CH₂)_x- OH, or-C (O)- OH；

M is integer 1 to 3；

N is selected from 1 to 3 integer；

X is 0 or integer 1 to 6.

2. formula (I) compound according to claim 1, wherein：

M is 1；

N is 2 or 3；And

R4 is-C (O) NRbRc, wherein R_bAnd R_cAs defined in claim 1.

3. formula (I) compound according to claim 1, wherein R¹It is selected from：

4. formula (I) compound according to claim 1, wherein R¹It is selected from：

5. formula (I) compound according to claim 1, wherein wherein R⁴It is selected from：

On the other hand, the present invention relates separately to formula (I) to (IV) compound, wherein R⁴It is selected from：

6. formula (III) compound or its pharmaceutically acceptable salt：

Wherein：

X₁Or X₂It is independently selected from-N or-C；

Wherein：

Work as X₂When being-N, R₂It is not present；Or

Work as X₂When being-C, R₂As defined above；

Each R⁵And R⁶It is independently selected from hydrogen, straight or branched C₁-C₆Alkyl, straight or branched-C₁-C₆Haloalkyl ,-C₁-C₆Cycloalkanes Base ,-(CH₂)_xC_1-C₆Cycloalkyl, heterocyclic radical ,-N- heterocyclic radicals, aryl, heteroaryl, or-(CH₂)_xHeteroaryl ,-(CHR_g)_xHeteroaryl Base；

Wherein：

Each R as defined above¹、R²、R³、R⁵And R⁶Optionally further it is selected from following one or more substituents substitution：Hydrogen, halogen Element ,-OH ,-(CH₂)_xOH、-C≡N、-NR_dR_e, straight or branched C₁-C₆Alkyl, straight or branched-C₁-C₆Haloalkyl, straight chain Or side chain C₁-C₆Alkoxy, straight or branched C₁-C₆Halogenated alkoxy ,-O- straight or brancheds-C₁-C₆Haloalkyl ,-C₁-C₆Ring Alkyl ,-(CH₂)_x- cycloalkyl, heterocyclic radical, aryl ,-heteroaryl ,-(CH₂)_x- heteroaryl ,-O- (CH₂)_xCH(OH)CH₂(OH), Or-C (O) OR_f；

Each R as defined above_a、R_b、R_c、R_d、R_e、R_fOr R_gIt is independently selected from hydrogen, straight or branched C₁-C₆Alkyl, straight or branched- C₁-C₆Haloalkyl ,-C₁-C₆- cycloalkyl ,-(CH₂)_xC₁-C₆- cycloalkyl, heterocyclic radical ,-N- heterocyclic radicals, aryl, heteroaryl, or- (CH₂)_xHeteroaryl；

Wherein：

Each R as defined above_a、R_b、R_c、R_d、R_e、R_fOr R_gOptionally further it is selected from following one or more substituents substitution： Hydrogen, halogen ,-OH ,-(CH₂)_xOH ,-C ≡ N ,-NR_hR_i, straight or branched C₁-C₆Alkyl, straight or branched-C₁-C₆Haloalkyl, Straight or branched C₁-C₆Alkoxy, straight or branched-C₁-C₆Halogenated alkoxy ,-C₁-C₆Cycloalkyl ,-(CH₂)_xIt is-cycloalkyl, miscellaneous Ring group ,-heterocyclic radical ,-O- heterocyclic radicals, aryl ,-heteroaryl ,-(CH₂)_x- heteroaryl ,-O- (CH₂)_xCH(OH)CH₂(OH)、- (CH₂)_x- OH or-C (O) OR_j；

Wherein：

M is integer 1 to 3；

N is selected from 2 to 3 integer；

X is 0 or selected from 1 to 6 integer.

7. formula (V) compound or its pharmaceutically acceptable salt:

Wherein：

Each R⁵And R⁶It is independently selected from hydrogen, straight or branched C₁-C₆Alkyl, straight or branched-C₁-C₆Haloalkyl ,-C₁-C₆Cycloalkanes Base ,-(CH₂)_xC₁-C₆Cycloalkyl, heterocyclic radical ,-N- heterocyclic radicals, aryl, heteroaryl, or-(CH₂)_xHeteroaryl ,-(CHR_g)_xHeteroaryl Base；

Wherein：

Each R as defined above_a、R_b、R_c、R_d、R_e、R_fOr R_gOptionally further it is selected from following one or more substituents substitution： Hydrogen, halogen ,-OH ,-(CH₂)_xOH ,-C ≡ N ,-NR_hR_i, straight or branched C₁-C₆Alkyl, straight or branched-C₁-C₆Haloalkyl, Straight or branched C₁-C₆Alkoxy, straight or branched-C₁-C₆Halogenated alkoxy ,-C₁-C₆Cycloalkyl ,-(CH₂)_xIt is-cycloalkyl, miscellaneous Ring group ,-heterocyclic radical ,-O- heterocyclic radicals, aryl ,-heteroaryl ,-(CH₂)_x- heteroaryl ,-O- (CH₂)_xCH(OH)CH₂(OH)、- (CH₂)_x- OH, or-C (O) OR_j；

Wherein：

M is integer 1 to 3；

N is selected from 2 to 3 integer；

X is 0 or selected from 1 to 6 integer.

8. compound, it is the compound listed in Table I or its pharmaceutically acceptable salt.

9. pharmaceutical composition, it includes the compound and pharmaceutically acceptable carrier of any one of claim 1 to 8.

10. the pharmaceutical composition of claim 9, also comprising extra activating agent.

11. for treating insulin resistance, metabolic syndrome, metabolic dysfunction, diabetes or their complication, or it is used for Increase the method for insulin sensitivity, including give to subject in need the compound of any one of claim 1 to 8 Or the pharmaceutical composition of any one of claim 9 or 10.

12. treat by SIRT1 express or activity reduce caused by disease or obstacle method, it is included to there is the tested of this needs Person gives any one of compound or its pharmaceutically acceptable salt or claim 9 or 10 of any one of claim 1 to 8 Pharmaceutical composition.

13. method according to claim 12, wherein disease or the obstacle caused by SIRT1 expression or activity reduction are selected from But be not limited to, aging or stress, diabetes, metabolic dysfunction, neurodegenerative disease, angiocardiopathy, cancer or inflammatory disease Disease.

14. method according to claim 13, wherein to aging or stress be related disease, diabetes, metabolic dysfunction, god Psoriasis, atopic dermatitis, acne, brandy nose, inflammatory bowel are selected from through degenerative disease, angiocardiopathy, cancer or inflammatory disease Disease, osteoporosis, septicemia, arthritis, COPD, systemic loupus erythematosus and ophthalmic inflammation.

15. treating the method for psoriasis, it includes giving the change of any one of claim 1 to 8 to subject in need The pharmaceutical composition of compound or claim 9 or 10.

16. the compound as defined in claim 1 to 8 compound, for treat suffer from or be susceptible to suffer from insulin resistance, be metabolized it is comprehensive In the therapy of the subject of simulator sickness diabetes or their complication, or for increasing insulin sensitivity in subject.

17. compound as defined in claim 1 to 8 compound prepare be used to treating in subject insulin resistance, Purposes in metabolic syndrome, diabetes or their complication or medicine for increasing insulin sensitivity.